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TAPESHWAR YADAV
(LECTURER)
BMLT, DNHE,
M.SC. MEDICAL
BIOCHEMISTRY
Clinical or Medical
Laboratory
A – A good student is liked by teacher
G – Greets everyone with smile
O – Obedient
O – On time for college
D – Dresses neatly
S – Studies with interest
T – Treats everyone with smile
U – Understands everything
D – Does daily home work
E – Eager to know new things
N – Never misbehaves
T – Talks little in class
Introduction
Laboratory is a place that is equipped with different
instruments, equipments and chemicals (reagents) etc., for
performing experimental works, research activities and
investigative procedures.
Medical laboratory is one part of the laboratory that is
equipped with various biomedical instruments,
equipments, materials and reagents (chemicals) for
performing different laboratory investigative activities by
using biological specimens (whole blood, serum, plasma,
urine, stool, etc).
Medical laboratory science
Medical laboratory science is a complex field
embracing a number of different disciplines
such as:
Microbiology,
Hematology,
Clinical Chemistry,
Urinalysis, Immunology, Serology,
Histopathology, Immunohematology and
Molecular biology and others.
Contd…
Immunology,
Serology,
Histopathology,
Immunohematology and
Molecular biology and others.
Medical Laboratory Technology
The practice of modern medicine would be
impossible without the tests performed in the
clinical laboratory.
 A medical team of pathologists, specialists,
scientists, technologists, and technicians work
together to determine the presence, extent, or
absence of disease and provide data needed to
evaluate the effectiveness of treatment.
Clinical or Medical Laboratory
Laboratories that perform chemical and microscopic
tests on:
blood
other body fluids
tissues
Clinical Laboratories
 Play a major role in patient care
 Variety of settings
 Two types of Clinical.Laboratory
 Hospital lab.
 Non hospital lab.
 POLs
 Reference laboratories(LABCORP/QUEST D.)
 Government laboratories - federal
 Center for Disease control and Prevention(CDC)
 Epidemiology labs
 Laboratory Response Network
 Government Laboratories- state
 Premarital blood testing
 PKU testing in newborns
 Fungi,virus, and mycobacteria culture
Regulations of Clinical Laboratory
 All laboratories, but research labs.are regulated by
Federal and State agencies
 CLIA’88- Clinical Laboratory Improvement
Amendments of 1988:
Is a revision to the original CLIA of 1967, specifies
the minimum performance standards for all
Clinical Laboratories
Objectives of CLIA’88
 To ensure quality Laboratory Testing, amendments
are continually revised, updated, clarified and
refined
 CMS:Center for Medicare and Medicaid
Services,agency within the Department of Health
and Human Services responsible for implementing
CLIA’88
CMS
 Any Laboratory performing Lab. tests in humans
,except for research Labs. Must obtain a certificate
from CMS (center for medicare-medicaid services) to
be allowed to operate
Laboratory Personnel
 Director of the Lab.- Pathologist, MD, DO, or hold
a doctorate in a related clinical field. Hold
certification and have supervisory and clinical
laboratory experience
 Technical supervisor/Lab.Manager-someone
educated in the clinical laboratory sciences who
has additional business experience
Laboratory personnel
 General supervisor for each area
 Testing personnel:
 Medical Technologists(MT/CLS)
 Medical Lab.Technicians(MLT/CLT)
 Medical assistants/nursing staff(POLs)
Departments of the Clinical Laboratory
 Clinical Chemistry
 Hematology
 Microbiology
 Blood Bank
 Supports Services (Phlebotomy/Specimen
Processing)
Clinical Chemistry
 Tests perform in serum, plasma, urine and other
body fluids such as spinal fluid, or joint fluid
 Largest department in the Lab.
 Toxicology
 Special chemistry
Hematology
 Studying of the cellular components of the blood
 Quantitative or Qualitative
 Coagulation
 Urinalysis
 Special hematology
Microbiology
 Culture/identification microorganisms
 From sputum, wounds, blood, urine and other body
fluids
 Inoculated in culture media
 Organisms are identified and susceptibility test are
performed
 Bacteriology, virology, serology, parasitology
Blood Bank
 Also called immunohematology or transfusion
services
 ABO group and Rh typing
 Antibody testing
 Storage of packed cells units
 Processing of some components like platelets and
cryoprecipitate
Support Services
 Phlebotomists
 Accessioning
POCT
 Point of care testing brings the laboratory to the
patient, also called bed-side testing
 Use small simple analyzers
 Portable instruments
 Hgb, glucose, electrolytes,and cholesterol
Quality Assessment System
 QA.is incorporated to each department’s procedure
manuals and day to day operation
 Standardized material are analyzed on each
instrument to document precision, and
reproducibility
 Calibration, maintenance and repair of the
instruments is recorded
 Participate in proficiency testing programs
 Health care agencies have very specific
standards, rules and regulations governing the
education and job responsibilities of the
laboratory personnel
 Lab. professionals are required to complete an
authorized program and certification
 Lab. Personnel need to observe/protect patient
privacy
Safety
 Occupational Safety and Health
Administration(OSHA) began in 1970 as a legislation
and subsequent rules that mandate increased
attention to safety in workplaces
 The Clinical laboratory has, physical, chemical and
biological hazards
PPE
 Employees in the clinical lab are required to use
personal protective equipment:
 Gloves
 Mask
 Gowns
Biohazards
 In 1980 clinical laboratory safety training
concentrated in protection from chemical,
physical,and contagious diseases such as
tuberculosis
 The discovery of AIDS, increased in Hepatitis B
virus(HBV) and Hepatitis C virus(HCV) brought an
emphasis on biological safety
 The term Biohazard came into use
 A Biohazard symbol was adopted that indicates the
presence of biological hazard or biohazardous
condition
Evolution on Biological safety
 By 1960 infectious patients were placed in
ISOLATION rooms
 1970-CDC outlined isolation guidelines and listed
isolation categories
 1985-in response to the increasing AIDS/HIV
epidemic CDC adopted Universal Blood and Body
fluids precautions, to be applied in all patients
regardless of their infectious status
 1987- Body substance Isolation, included all body
fluid even if not visibly contaminated with blood
Evolution on Biological safety
 1991-OSHA issued “Bloodborne pathogens
standard”, not included on previous regulation
 1996- CDC implemented “Standard Precautions”
that includes a comprehensive set of safety
guidelines for Health care workers rendering care
to patients, this is the current terminology
 To control nosocomial (inst. acquired) infections
 Transmission-based precautions(additional practices for
pathogens that spread by air, droplets, and contact
2001-OSHA revised the BBP(blood borne pathogen)
standard to prevent accidental needle-sticks in the
workplace
Standard Precautions
Requires that every patient and every body
fluid, body substance, organ, or unfixed tissue
be regarded as potentially infectious
 Hands wash(plain soap)
 After touching body fluids and contaminated items, after
removing gloves and between patient contact
 Wear gloves
 When touching blood/body fluids/secretions, wear clean
gloves when touching mucous membranes and nonintact
skin
 Wear mask/eye protection/face shield
 Activities that could generate splashes, spray of blood, body
fluids , or secretions
Standard Precautions, cont.
Patient care equipment
 should be handled to prevent transfer of
microorganisms to other patients and environment
Linen
 Handle, transport,and process in a manner to avoid
contamination of clothing and other patients or
environment
Occupational health and blood-borne pathogens
 Prevent injuries when using, handling, cleaning and
disposing sharps
 NEVER RECAP A USED NEEDLE
 Do not removed used needle from syringe by hand
 Disposed used sharps on puncture resistant containers
Standard Precautions,cont.
 Use resuscitation devices as an alternative to mouth to
mouth resuscitation
 Patient placement
 Use a private room for patients who can be a source of
contamination or patients who are not expected to
maintain hygiene or environmental control
 Environmental control
 Follow hospital procedures for routine care and
cleaning/desinfection of any soiled device, equipment
or environmental surface
General laboratory equipment
 Centrifuges- spin samples at high speeds
forcing the heavier particles to the bottom of
the container,e.g..separating plasma and blood
cells
 Safety tips
 Use Standard Precautions/PPE
 Load must be balanced
 Tubes must be capped during operation
 Do not open the centrifuge while rotor is moving
 Clean spills immediately with surface disinfectants
General laboratory equipment
Autoclaves- use steam under pressure to sterilize
medical/surgical instruments, or contaminated
materials before disposal
 Never open unless the chamber pressure reads zero
 Use heat-proof gloves to remove items
 When sterilizing liquids use loosely capped, heat resistant
containers, no more than half full
 Use an autoclave tray to prevent liquids from spilling
General laboratory equipment
 Laboratory balances
 Used to measure chemicals
 Use PPE and chemical safety precautions
 Be gentle, Balances are delicate equipment
General laboratory equipment
 Other equipments
 Refrigerators
 Water baths
 PH meters
 Incubators
 Thermometers
 freezer
The Microscope
 Is a delicate and expensive instrument , special
care must be taken in its use
 Various types of microscopes, two categories based
on type of illumination
 Light microscopes
 Bright-field- stained specimens
 Phase-contrast-unstained cells,urine sediment
 Epi-fluorescence microscope,specimens treated with fluorescent
dyes, syphilis, mycobacteria
 Electron microscopes:provides greater magnification in
medical research
Light microscope images
A-stained cell seen with bright field microscope
B-phase contrast image
C-epi-fluorescence microscopy,Borrelia burgdorferi
Parts of the Microscope
Parts of the Microscope
Oculars: monocular or binocular
Objective lenses: attached to the revolving nose
piece, at least 3 present: low, high dry, and oil
immersion lenses
Light condenser which focuses and directs light to
the objectives, iris diaphragm that regulates the
amount of light that strikes the object observed
Field diaphragm:help align the light
Coarse and fine adjustments:focusing knobs
Stage:support for the object been viewed
Microscope safety
 Safety
 observe electrical safety rules
 Glass slide handle with care to avoid breaking
 Unfixed specimens should be treated with standard
precautions,disinfect stage after use
 QA
 Scheduled maintenance should be performed and
documented
 Care and cleaning of lenses
 Use only lens paper, clean lenses before and after each use
 Do not allowed immersion oil to touch the low and high dry
lenses
 Transporting and storing
Transporting the Microscope
Using the Microscope
 Use low power objective to locate and to view large
objects
 With the coarse adjustment knob bring the objective
and the slide as close together as possible
 While looking through the oculars, move the coarse
adjustment knob to bring the objective and slide apart
until the object on the slide comes into focus
 Use the fine adj.knob to bring the image into sharp
focus
Using the Microscope
 If you need to use the high power(40x), to see cells
and sediments, after initial focusing with the low
power(20x), rotate the high power into position
 Never use the coarse adjustment knob with high
power, the distance between the objective and slide is
very small and the slide could break.
 Oil immersion lenses(100x) give the highest
magnification of the bright field objectives
Using oil immersion lenses
 After initially focusing with the low power, rotate
the objective to the side and place a small drop of
immersion oil on the slide
 The oil immersion objective is rotated into the drop
of oil been careful no other objective touch the
oil
 use only fine adjustment knob with oil
 Condenser should be all the way up
 Maximum light source
 Open the iris diaphragm to the maximum
After using the Microscope
 Always switch to the low magnification objective
 With lens paper clean the oil immersion objective,
stage and condenser if oil has become in contact
with it
 Turn the light source off
 Unplug the microscope
 Store in proper location or cover as appropriate
Calculate Magnification
 Degree of magnification on the ocular multiplied
by the degree of magnification on the objectives
Example:
10x(ocular) x 100x(oil immersion)= 1000x
The object viewed would be magnified
1000 times its original size
Resolving power: the ability of a microscope to
produce separate images of closely spaced details
in the object being viewed
Blood collection
 Capillary puncture: small amount of blood collected for
glucose, K, electrolytes, Hgb, Htc, Plt count, or when a
larger sample is difficult to obtain as in newborns
 Routine venipuncture: most common method of
obtaining blood, a superficial vein is punctured with a
hypodermic needle and blood is collected into a syringe
or vacuum tube
Capillary Puncture
 Safe
 Quick
 Small amount of blood
 Increased use
 Point-of-care testing (POCT)
 Physician Office Laboratories
Capillary Puncture Sites
 Fingertip
 Great toe
 Heel
Capillary Puncture Sites
Lancets
 Sterile
 Single-use
 Different lengths
Collection Containers
Procedure
Routine Venipuncture
 Phlebotomy
 Superficial vein
 Large sample of blood
 Skill and experience
 Preserve vein integrity
Venipuncture Supplies
 Needles
 Various safety designs
 21 ga, 1 inch
 Needle holders
 Phlebotomy tray
Venipuncture Supplies
Venipuncture Supplies
 Vacuum tubes and anticoagulants
 Sizes
 Stopper color:
 Red: no anticoagulant, to collect serum for blood chemistries
and serology tests
 Lavender: containing EDTA for hematologycal and blood
typing tests(ethylenediaminetetraacetic acid )
 Green: contains heparin, for lymphocytes studies and special
chemistry
 Light blue: sodium citrate for coagulation studies
 Gray :potasium oxalate, for glucose and legal alcohol
 Black: for westergren ESR
 Draw exact amount
Safety Precautions
 Observe standard precautions
 Wear gloves and other PPE
 Never recap needles
 Use proper technique
 Avoid
 Hemoconcentration: do not leave tourniquet in place for
more than 1-2 minutes
 Hemolysis: do not shake tubes, mix by gently inverting a
few times
Select Equipment
Patient Preparation
 Patient I.D.
 Explain procedure
 Support patient and arm
 Be prepared! for any sudden reaction from the
patient, or occasional patient who may faint
Patient Preparation
Apply Tourniquet
•3-4 inches above
elbow
•Use quick release
tie
Identify Suitable Vein
 Veins commonly used
 Median cubital
 Basilic
 Cephalic
 Palpate vein:
carefully inspect
both arms to find
the better site
Perform Venipuncture
 Alcohol-cleanse site, let air dry, do not touch the
site after cleaning
 Observe bevel up
 Anchor vein with thumb 1inch below the
puncture site
 Enter vein in the same direction of it, in a15-25
degree angle, in a smooth motion
 Insert vacuum tube
 Clot tube first
 Invert anticoagulant tubes softly 5-7 times
Perform Venipuncture
Adverse situations
 In case of patient developing a large hematoma while
venipuncture procedure is being done, withdraw the
needle, apply pressure, and intent the procedure in a
different site
 In case of failure to obtain the blood, ask the patient
permission for a second intent, if he agrees try in a
different site
 After the second non-productive intent,inform the
patient and find another person to draw the specimen
Complete Procedure
 Activate safety feature
 Immediate disposal
 Label tubes before leaving the room
 Patient care
Patient care
 The tourniquet is always release before needle is
withdraw
 Gauze should be applied over the puncture site and
pressure maintained for 1-3 minutes or until
bleeding stops
 Ask patient to keep arm extended
 Offer a small bandage if necessary
In Case of Accident
 Immediately clean exposed area
 Flood with water
 Clean with antiseptic soap
 Report immediately to supervisor
 Seek medical attention
Label the samples
 Must contain patient information
 Name
 Date of birth
 Date and time of collection
 And initials of the person drawing the blood
 Tubes should never be prelabeled to avoid using the
prelabeled tube in the wrong patient
 Make sure the tubes are clean and no blood has
contaminated the outer part of the tubes
 Place specimen in a biohazard labeled bag and proceed as
required by the institution
selecting tests to use:
 Test selections are based on :
 subjective clinical judgment,
 national recommendations,
 and evidence-based health care.
 Often diagnostic tests or procedures are used as
predictors of surgical risk or morbidity and mortality
rates because, in some cases, the risk may outweigh the
benefit.
selecting tests to use:
1.Basic screening (frequently used with wellness
groups and case finding)
2. Establishing (initial) diagnoses
3. Differential diagnosis
4. Evaluating current medical case management
and outcomes
5. Evaluating disease severity
6. Monitoring course of illness and response to
treatment
7. Group and panel testing
8. Regularly scheduled screening tests as part of
ongoing care
9. Testing related to specific events, certain signs and
symptoms, or other exceptional situations (eg, infection
and inflammation , sexual assault, drug screening,
postmortem tests, to name a few)
groups and case finding)
 Cervical Papanicolaou (Pap) test
 Yearly for all women 18 years of age; more often with
high-risk factors (eg, dysplasia, human
immunodeficiency virus [HIV], herpes simplex);
check for human papillomavirus (HPV), chlamydia,
and gonorrhea using DNA
Establishing (initial) diagnoses
 Serum amylase
 In the presence of abdominal pain, suspect pancreatitis
 Thyroid-stimulating hormone (TSH) test
 Suspicion of hypothyroidism, hyperthyroidism, or thyroid
dysfunction in patients 50 years of age
Differential diagnosis
 Chlamydia and gonorrhea
 In sexually active persons with multiple partners; monitor for
pelvic inflammatory disease
Evaluating current medical case
management and outcomes
 Tuberculosis (TB) blood test QuantiFERON Gold TB
 Blood test to assess TB exposure in risk population
 Syphilis serum fluorescent treponemal antibody
(FTA) test
 Positive rapid plasma reagin (RPR) test result
Grading Guidelines for Scientific
Evidence
 A. Clear evidence from all appropriately conducted
trials
 Measure plasma glucose through an accredited lab to diagnose or
screen for diabetes
 B.Supportive evidence from well-conducted studies or
registries
 Draw fasting blood plasma specimens for glucose analysis
 C.No published evidence; or only case,
observational, or historical evidence •
 Self-monitoring of blood glucose may help to achieve better
control
 E.Expert consensus or clinical experience or
Internet polls
 Measure ketones in urine or blood to monitor and diagnose
diabetic ketoacidosis (DKA) (in home or clinic)
The diagnostic testing model
 incorporates three phases:
 pretest,
 emphasis on appropriate test selection,
 obtaining proper consent,
 proper patient preparation,
 individualized patient education,
 emotional support, and effective communication.
 These interventions are key to achieving the desired outcomes
and preventing misunderstandings and errors.
 Intratest Phase: Elements of Safe, Effective,
Informed Care
 Posttest Phase: Elements of Safe, Effective,
Informed Care
The clinical value of a test is related to
 sensitivity, specificity, and the incidence of the
disease in the population tested.
 Sensitivity and specificity do not change with
different populations of ill and healthy patients
 The predictive value of the same test can vary
significantly with age, gender, and geographic
location.
 Specificity refers to the ability of a test to identify
correctly those individuals who do not have the
disease.
 The division formula for specificity is as follows:
 Specificity%=persons w/o dis.who test neg./total #
of persons w/o dis. X 100
 Sensitivity refers to the ability of a test to
correctly identify those individuals who truly
have the disease.
 The division formula for sensitivity is as follows:
 Sensitivity% = persons with dis.who test
positive/ total # persons tested with disease x
100
 Incidence refers to the number of new cases of a
disease, during a specified period of time, in a
specified population or community.
 Prevalence refers to the number of existing cases
of a disease, at a specific period of time, in a
given population.
Predictive values
Predictive values refer to the ability of a screening
test result to correctly identify the disease state.
The predictive value of the same test can be very
different when applied to people of differing ages,
gender, geographic locations, and cultures.
test outcome deviations
Minimize test outcome deviations
 following proper test protocols.
 Make certain the patient and his or her significant others know
what is expected of them.
 Written instructions are very helpful.
Reasons for deviations may include the
following
 Incorrect specimen collection, handling, storage, or
labeling
 Wrong preservative or lack of preservative
 Delayed specimen deliver
Reasons for deviations may include the
following
 Incorrect or incomplete patient preparation
 Hemolyzed blood samples
 Incomplete sample collection, especially of timed
samples
 Old or deteriorating specimens
Patient factors that can alter test results
may include the following
 Incorrect pretest diet
 Current drug therapy
 Type of illness.
 Dehydration
 Position or activity at time of specimen collection
Patient factors that can alter test results
may include the following
 Postprandial status (ie, time patient last ate)
 Time of day
 Pregnancy
 Age and Gender
Patient factors that can alter test results
may include the following
 Level of patient knowledge and understanding of
testing process
 Stress
 Nonadherence or noncompliance with
instructions and pretest preparation
 Undisclosed drug or alcohol use
avoid costly mistakes
 Communication errors account for more incorrect
results than do technical errors.
 Properly identify and label every specimen as soon as
it is obtained.
Educate the patient and family
 Educate regarding the testing process and what will
be expected
 Record the date, time, type of teaching, information
given, and person to whom the information was
given.
Educate the patient and family
 Giving sensory and objective information that relates
to what the patient will likely physically feel and the
equipment that will be used is important so that
patients can envision a realistic representation of
what will occur.
Educate the patient and family
 Avoid technical and medical jargon
 and adapt information to the patient's level of
understanding.
 Slang terms may be necessary to get a point across.
Educate the patient and family
 Encourage questions and verbalization of feelings,
fears, and concerns
 Do not dismiss, minimize, or invalidate the patient's
anxiety
 Develop listening skills, and be aware of nonverbal
signals (ie, body language)
Educate the patient and family
 Above all, be nonjudgmental.
 Emphasize that there is usually a waiting period (ie,
turn-around time) before test results are relayed
back to the clinicians and nursing unit.
 Offer listening, presence, and support during this
time of great concern and anxiety
Educate the patient and family
 Because of factors such as anxiety, language barriers,
and physical or emotional impairments, the patient
may not fully understand and assimilate instructions
and explanations
Educate the patient and family
 To validate the patient's understanding of what is
presented, ask the patient to repeat instructions
given to evaluate assimilation and understanding of
presented information.
normal or reference values
 Normal values are those that fall within 2 standard
deviations (ie, random variation) of the mean value
for the normal population.
 Normal ranges can vary to some degree from
laboratory to laboratory. Frequently, this is because
of the particular type of equipment used
normal or reference values
 The reported reference range for a test can vary
according to the laboratory used, the method
employed, the population tested, and methods of
specimen collection and preservation.
normal or reference values
 Interpretation of laboratory results must always be
in the context of the patient's state of being.
 Circumstances such as hydration, nutrition, fasting
state, mental status, or compliance with test
protocols are only a few of the situations that can
influence test outcomes.
clinical laboratory data values
 may be reported in conventional units, SI
units(Systéme International (SI) units), or both
 The SI system uses seven dimensionally independent
units of measurement to provide logical and
consistent measurements
clinical laboratory data values
SI concentrations are written as amount per volume
 (moles or millimoles per liter)
 rather than as mass per volume (grams, milligrams,
or milliequivalents per deciliter, 100 milliliters, or
liter)
 Numerical values may differ between systems
or may be the same.
 For example, chloride is the same in both
systems: 95 to 105 mEq/L (conventional)
 and 95 to 105 mmol/L (SI).
Recognize margins of error
 possibility exists that some tests will be abnormal owing
purely to chance
 because a significant margin of error arises from the
arbitrary setting of limits.
 Moreover, if a laboratory test is considered normal up to
the 95th percentile, then 5 times out of 100, the test will
show an abnormality even though a patient is not ill
Cultural Sensitivity
 Many cultures have diverse beliefs about diagnostic
testing that requires blood sampling
 Preserving the cultural well-being of any individual
or group promotes compliance with testing and
easier recovery from routine as well as more invasive
and complex procedures
END

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TAPESHWAR YADAV'S GUIDE TO MEDICAL LABORATORY SCIENCE

  • 1. TAPESHWAR YADAV (LECTURER) BMLT, DNHE, M.SC. MEDICAL BIOCHEMISTRY Clinical or Medical Laboratory
  • 2. A – A good student is liked by teacher G – Greets everyone with smile O – Obedient O – On time for college D – Dresses neatly S – Studies with interest T – Treats everyone with smile U – Understands everything D – Does daily home work E – Eager to know new things N – Never misbehaves T – Talks little in class
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  • 4. Introduction Laboratory is a place that is equipped with different instruments, equipments and chemicals (reagents) etc., for performing experimental works, research activities and investigative procedures. Medical laboratory is one part of the laboratory that is equipped with various biomedical instruments, equipments, materials and reagents (chemicals) for performing different laboratory investigative activities by using biological specimens (whole blood, serum, plasma, urine, stool, etc).
  • 5. Medical laboratory science Medical laboratory science is a complex field embracing a number of different disciplines such as: Microbiology, Hematology, Clinical Chemistry, Urinalysis, Immunology, Serology, Histopathology, Immunohematology and Molecular biology and others.
  • 7. Medical Laboratory Technology The practice of modern medicine would be impossible without the tests performed in the clinical laboratory.  A medical team of pathologists, specialists, scientists, technologists, and technicians work together to determine the presence, extent, or absence of disease and provide data needed to evaluate the effectiveness of treatment.
  • 8. Clinical or Medical Laboratory Laboratories that perform chemical and microscopic tests on: blood other body fluids tissues
  • 9. Clinical Laboratories  Play a major role in patient care  Variety of settings  Two types of Clinical.Laboratory  Hospital lab.  Non hospital lab.  POLs  Reference laboratories(LABCORP/QUEST D.)  Government laboratories - federal  Center for Disease control and Prevention(CDC)  Epidemiology labs  Laboratory Response Network
  • 10.  Government Laboratories- state  Premarital blood testing  PKU testing in newborns  Fungi,virus, and mycobacteria culture
  • 11. Regulations of Clinical Laboratory  All laboratories, but research labs.are regulated by Federal and State agencies  CLIA’88- Clinical Laboratory Improvement Amendments of 1988: Is a revision to the original CLIA of 1967, specifies the minimum performance standards for all Clinical Laboratories
  • 12. Objectives of CLIA’88  To ensure quality Laboratory Testing, amendments are continually revised, updated, clarified and refined  CMS:Center for Medicare and Medicaid Services,agency within the Department of Health and Human Services responsible for implementing CLIA’88
  • 13. CMS  Any Laboratory performing Lab. tests in humans ,except for research Labs. Must obtain a certificate from CMS (center for medicare-medicaid services) to be allowed to operate
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  • 15. Laboratory Personnel  Director of the Lab.- Pathologist, MD, DO, or hold a doctorate in a related clinical field. Hold certification and have supervisory and clinical laboratory experience  Technical supervisor/Lab.Manager-someone educated in the clinical laboratory sciences who has additional business experience
  • 16. Laboratory personnel  General supervisor for each area  Testing personnel:  Medical Technologists(MT/CLS)  Medical Lab.Technicians(MLT/CLT)  Medical assistants/nursing staff(POLs)
  • 17. Departments of the Clinical Laboratory  Clinical Chemistry  Hematology  Microbiology  Blood Bank  Supports Services (Phlebotomy/Specimen Processing)
  • 18. Clinical Chemistry  Tests perform in serum, plasma, urine and other body fluids such as spinal fluid, or joint fluid  Largest department in the Lab.  Toxicology  Special chemistry
  • 19. Hematology  Studying of the cellular components of the blood  Quantitative or Qualitative  Coagulation  Urinalysis  Special hematology
  • 20. Microbiology  Culture/identification microorganisms  From sputum, wounds, blood, urine and other body fluids  Inoculated in culture media  Organisms are identified and susceptibility test are performed  Bacteriology, virology, serology, parasitology
  • 21. Blood Bank  Also called immunohematology or transfusion services  ABO group and Rh typing  Antibody testing  Storage of packed cells units  Processing of some components like platelets and cryoprecipitate
  • 23. POCT  Point of care testing brings the laboratory to the patient, also called bed-side testing  Use small simple analyzers  Portable instruments  Hgb, glucose, electrolytes,and cholesterol
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  • 25. Quality Assessment System  QA.is incorporated to each department’s procedure manuals and day to day operation  Standardized material are analyzed on each instrument to document precision, and reproducibility  Calibration, maintenance and repair of the instruments is recorded  Participate in proficiency testing programs
  • 26.  Health care agencies have very specific standards, rules and regulations governing the education and job responsibilities of the laboratory personnel  Lab. professionals are required to complete an authorized program and certification  Lab. Personnel need to observe/protect patient privacy
  • 27. Safety  Occupational Safety and Health Administration(OSHA) began in 1970 as a legislation and subsequent rules that mandate increased attention to safety in workplaces  The Clinical laboratory has, physical, chemical and biological hazards
  • 28. PPE  Employees in the clinical lab are required to use personal protective equipment:  Gloves  Mask  Gowns
  • 29. Biohazards  In 1980 clinical laboratory safety training concentrated in protection from chemical, physical,and contagious diseases such as tuberculosis  The discovery of AIDS, increased in Hepatitis B virus(HBV) and Hepatitis C virus(HCV) brought an emphasis on biological safety  The term Biohazard came into use  A Biohazard symbol was adopted that indicates the presence of biological hazard or biohazardous condition
  • 30. Evolution on Biological safety  By 1960 infectious patients were placed in ISOLATION rooms  1970-CDC outlined isolation guidelines and listed isolation categories  1985-in response to the increasing AIDS/HIV epidemic CDC adopted Universal Blood and Body fluids precautions, to be applied in all patients regardless of their infectious status  1987- Body substance Isolation, included all body fluid even if not visibly contaminated with blood
  • 31. Evolution on Biological safety  1991-OSHA issued “Bloodborne pathogens standard”, not included on previous regulation  1996- CDC implemented “Standard Precautions” that includes a comprehensive set of safety guidelines for Health care workers rendering care to patients, this is the current terminology  To control nosocomial (inst. acquired) infections  Transmission-based precautions(additional practices for pathogens that spread by air, droplets, and contact 2001-OSHA revised the BBP(blood borne pathogen) standard to prevent accidental needle-sticks in the workplace
  • 32. Standard Precautions Requires that every patient and every body fluid, body substance, organ, or unfixed tissue be regarded as potentially infectious  Hands wash(plain soap)  After touching body fluids and contaminated items, after removing gloves and between patient contact  Wear gloves  When touching blood/body fluids/secretions, wear clean gloves when touching mucous membranes and nonintact skin  Wear mask/eye protection/face shield  Activities that could generate splashes, spray of blood, body fluids , or secretions
  • 33. Standard Precautions, cont. Patient care equipment  should be handled to prevent transfer of microorganisms to other patients and environment Linen  Handle, transport,and process in a manner to avoid contamination of clothing and other patients or environment Occupational health and blood-borne pathogens  Prevent injuries when using, handling, cleaning and disposing sharps  NEVER RECAP A USED NEEDLE  Do not removed used needle from syringe by hand  Disposed used sharps on puncture resistant containers
  • 34. Standard Precautions,cont.  Use resuscitation devices as an alternative to mouth to mouth resuscitation  Patient placement  Use a private room for patients who can be a source of contamination or patients who are not expected to maintain hygiene or environmental control  Environmental control  Follow hospital procedures for routine care and cleaning/desinfection of any soiled device, equipment or environmental surface
  • 35. General laboratory equipment  Centrifuges- spin samples at high speeds forcing the heavier particles to the bottom of the container,e.g..separating plasma and blood cells  Safety tips  Use Standard Precautions/PPE  Load must be balanced  Tubes must be capped during operation  Do not open the centrifuge while rotor is moving  Clean spills immediately with surface disinfectants
  • 36. General laboratory equipment Autoclaves- use steam under pressure to sterilize medical/surgical instruments, or contaminated materials before disposal  Never open unless the chamber pressure reads zero  Use heat-proof gloves to remove items  When sterilizing liquids use loosely capped, heat resistant containers, no more than half full  Use an autoclave tray to prevent liquids from spilling
  • 37. General laboratory equipment  Laboratory balances  Used to measure chemicals  Use PPE and chemical safety precautions  Be gentle, Balances are delicate equipment
  • 38. General laboratory equipment  Other equipments  Refrigerators  Water baths  PH meters  Incubators  Thermometers  freezer
  • 39. The Microscope  Is a delicate and expensive instrument , special care must be taken in its use  Various types of microscopes, two categories based on type of illumination  Light microscopes  Bright-field- stained specimens  Phase-contrast-unstained cells,urine sediment  Epi-fluorescence microscope,specimens treated with fluorescent dyes, syphilis, mycobacteria  Electron microscopes:provides greater magnification in medical research
  • 40. Light microscope images A-stained cell seen with bright field microscope B-phase contrast image C-epi-fluorescence microscopy,Borrelia burgdorferi
  • 41. Parts of the Microscope
  • 42. Parts of the Microscope Oculars: monocular or binocular Objective lenses: attached to the revolving nose piece, at least 3 present: low, high dry, and oil immersion lenses Light condenser which focuses and directs light to the objectives, iris diaphragm that regulates the amount of light that strikes the object observed Field diaphragm:help align the light Coarse and fine adjustments:focusing knobs Stage:support for the object been viewed
  • 43. Microscope safety  Safety  observe electrical safety rules  Glass slide handle with care to avoid breaking  Unfixed specimens should be treated with standard precautions,disinfect stage after use  QA  Scheduled maintenance should be performed and documented  Care and cleaning of lenses  Use only lens paper, clean lenses before and after each use  Do not allowed immersion oil to touch the low and high dry lenses  Transporting and storing
  • 45. Using the Microscope  Use low power objective to locate and to view large objects  With the coarse adjustment knob bring the objective and the slide as close together as possible  While looking through the oculars, move the coarse adjustment knob to bring the objective and slide apart until the object on the slide comes into focus  Use the fine adj.knob to bring the image into sharp focus
  • 46. Using the Microscope  If you need to use the high power(40x), to see cells and sediments, after initial focusing with the low power(20x), rotate the high power into position  Never use the coarse adjustment knob with high power, the distance between the objective and slide is very small and the slide could break.  Oil immersion lenses(100x) give the highest magnification of the bright field objectives
  • 47. Using oil immersion lenses  After initially focusing with the low power, rotate the objective to the side and place a small drop of immersion oil on the slide  The oil immersion objective is rotated into the drop of oil been careful no other objective touch the oil  use only fine adjustment knob with oil  Condenser should be all the way up  Maximum light source  Open the iris diaphragm to the maximum
  • 48. After using the Microscope  Always switch to the low magnification objective  With lens paper clean the oil immersion objective, stage and condenser if oil has become in contact with it  Turn the light source off  Unplug the microscope  Store in proper location or cover as appropriate
  • 49. Calculate Magnification  Degree of magnification on the ocular multiplied by the degree of magnification on the objectives Example: 10x(ocular) x 100x(oil immersion)= 1000x The object viewed would be magnified 1000 times its original size Resolving power: the ability of a microscope to produce separate images of closely spaced details in the object being viewed
  • 50. Blood collection  Capillary puncture: small amount of blood collected for glucose, K, electrolytes, Hgb, Htc, Plt count, or when a larger sample is difficult to obtain as in newborns  Routine venipuncture: most common method of obtaining blood, a superficial vein is punctured with a hypodermic needle and blood is collected into a syringe or vacuum tube
  • 51. Capillary Puncture  Safe  Quick  Small amount of blood  Increased use  Point-of-care testing (POCT)  Physician Office Laboratories
  • 52. Capillary Puncture Sites  Fingertip  Great toe  Heel
  • 57. Routine Venipuncture  Phlebotomy  Superficial vein  Large sample of blood  Skill and experience  Preserve vein integrity
  • 58. Venipuncture Supplies  Needles  Various safety designs  21 ga, 1 inch  Needle holders  Phlebotomy tray
  • 60. Venipuncture Supplies  Vacuum tubes and anticoagulants  Sizes  Stopper color:  Red: no anticoagulant, to collect serum for blood chemistries and serology tests  Lavender: containing EDTA for hematologycal and blood typing tests(ethylenediaminetetraacetic acid )  Green: contains heparin, for lymphocytes studies and special chemistry  Light blue: sodium citrate for coagulation studies  Gray :potasium oxalate, for glucose and legal alcohol  Black: for westergren ESR  Draw exact amount
  • 61. Safety Precautions  Observe standard precautions  Wear gloves and other PPE  Never recap needles  Use proper technique  Avoid  Hemoconcentration: do not leave tourniquet in place for more than 1-2 minutes  Hemolysis: do not shake tubes, mix by gently inverting a few times
  • 63. Patient Preparation  Patient I.D.  Explain procedure  Support patient and arm  Be prepared! for any sudden reaction from the patient, or occasional patient who may faint
  • 65. Apply Tourniquet •3-4 inches above elbow •Use quick release tie
  • 66. Identify Suitable Vein  Veins commonly used  Median cubital  Basilic  Cephalic  Palpate vein: carefully inspect both arms to find the better site
  • 67. Perform Venipuncture  Alcohol-cleanse site, let air dry, do not touch the site after cleaning  Observe bevel up  Anchor vein with thumb 1inch below the puncture site  Enter vein in the same direction of it, in a15-25 degree angle, in a smooth motion  Insert vacuum tube  Clot tube first  Invert anticoagulant tubes softly 5-7 times
  • 69. Adverse situations  In case of patient developing a large hematoma while venipuncture procedure is being done, withdraw the needle, apply pressure, and intent the procedure in a different site  In case of failure to obtain the blood, ask the patient permission for a second intent, if he agrees try in a different site  After the second non-productive intent,inform the patient and find another person to draw the specimen
  • 70. Complete Procedure  Activate safety feature  Immediate disposal  Label tubes before leaving the room  Patient care
  • 71. Patient care  The tourniquet is always release before needle is withdraw  Gauze should be applied over the puncture site and pressure maintained for 1-3 minutes or until bleeding stops  Ask patient to keep arm extended  Offer a small bandage if necessary
  • 72. In Case of Accident  Immediately clean exposed area  Flood with water  Clean with antiseptic soap  Report immediately to supervisor  Seek medical attention
  • 73. Label the samples  Must contain patient information  Name  Date of birth  Date and time of collection  And initials of the person drawing the blood  Tubes should never be prelabeled to avoid using the prelabeled tube in the wrong patient  Make sure the tubes are clean and no blood has contaminated the outer part of the tubes  Place specimen in a biohazard labeled bag and proceed as required by the institution
  • 74. selecting tests to use:  Test selections are based on :  subjective clinical judgment,  national recommendations,  and evidence-based health care.  Often diagnostic tests or procedures are used as predictors of surgical risk or morbidity and mortality rates because, in some cases, the risk may outweigh the benefit.
  • 75. selecting tests to use: 1.Basic screening (frequently used with wellness groups and case finding) 2. Establishing (initial) diagnoses 3. Differential diagnosis 4. Evaluating current medical case management and outcomes 5. Evaluating disease severity
  • 76. 6. Monitoring course of illness and response to treatment 7. Group and panel testing 8. Regularly scheduled screening tests as part of ongoing care 9. Testing related to specific events, certain signs and symptoms, or other exceptional situations (eg, infection and inflammation , sexual assault, drug screening, postmortem tests, to name a few)
  • 77. groups and case finding)  Cervical Papanicolaou (Pap) test  Yearly for all women 18 years of age; more often with high-risk factors (eg, dysplasia, human immunodeficiency virus [HIV], herpes simplex); check for human papillomavirus (HPV), chlamydia, and gonorrhea using DNA
  • 78. Establishing (initial) diagnoses  Serum amylase  In the presence of abdominal pain, suspect pancreatitis  Thyroid-stimulating hormone (TSH) test  Suspicion of hypothyroidism, hyperthyroidism, or thyroid dysfunction in patients 50 years of age
  • 79. Differential diagnosis  Chlamydia and gonorrhea  In sexually active persons with multiple partners; monitor for pelvic inflammatory disease
  • 80. Evaluating current medical case management and outcomes  Tuberculosis (TB) blood test QuantiFERON Gold TB  Blood test to assess TB exposure in risk population  Syphilis serum fluorescent treponemal antibody (FTA) test  Positive rapid plasma reagin (RPR) test result
  • 81. Grading Guidelines for Scientific Evidence  A. Clear evidence from all appropriately conducted trials  Measure plasma glucose through an accredited lab to diagnose or screen for diabetes  B.Supportive evidence from well-conducted studies or registries  Draw fasting blood plasma specimens for glucose analysis
  • 82.  C.No published evidence; or only case, observational, or historical evidence •  Self-monitoring of blood glucose may help to achieve better control  E.Expert consensus or clinical experience or Internet polls  Measure ketones in urine or blood to monitor and diagnose diabetic ketoacidosis (DKA) (in home or clinic)
  • 83. The diagnostic testing model  incorporates three phases:  pretest,  emphasis on appropriate test selection,  obtaining proper consent,  proper patient preparation,  individualized patient education,  emotional support, and effective communication.  These interventions are key to achieving the desired outcomes and preventing misunderstandings and errors.
  • 84.  Intratest Phase: Elements of Safe, Effective, Informed Care  Posttest Phase: Elements of Safe, Effective, Informed Care
  • 85. The clinical value of a test is related to  sensitivity, specificity, and the incidence of the disease in the population tested.  Sensitivity and specificity do not change with different populations of ill and healthy patients  The predictive value of the same test can vary significantly with age, gender, and geographic location.
  • 86.  Specificity refers to the ability of a test to identify correctly those individuals who do not have the disease.  The division formula for specificity is as follows:  Specificity%=persons w/o dis.who test neg./total # of persons w/o dis. X 100
  • 87.  Sensitivity refers to the ability of a test to correctly identify those individuals who truly have the disease.  The division formula for sensitivity is as follows:  Sensitivity% = persons with dis.who test positive/ total # persons tested with disease x 100
  • 88.  Incidence refers to the number of new cases of a disease, during a specified period of time, in a specified population or community.  Prevalence refers to the number of existing cases of a disease, at a specific period of time, in a given population.
  • 89. Predictive values Predictive values refer to the ability of a screening test result to correctly identify the disease state. The predictive value of the same test can be very different when applied to people of differing ages, gender, geographic locations, and cultures.
  • 90. test outcome deviations Minimize test outcome deviations  following proper test protocols.  Make certain the patient and his or her significant others know what is expected of them.  Written instructions are very helpful.
  • 91. Reasons for deviations may include the following  Incorrect specimen collection, handling, storage, or labeling  Wrong preservative or lack of preservative  Delayed specimen deliver
  • 92. Reasons for deviations may include the following  Incorrect or incomplete patient preparation  Hemolyzed blood samples  Incomplete sample collection, especially of timed samples  Old or deteriorating specimens
  • 93. Patient factors that can alter test results may include the following  Incorrect pretest diet  Current drug therapy  Type of illness.  Dehydration  Position or activity at time of specimen collection
  • 94. Patient factors that can alter test results may include the following  Postprandial status (ie, time patient last ate)  Time of day  Pregnancy  Age and Gender
  • 95. Patient factors that can alter test results may include the following  Level of patient knowledge and understanding of testing process  Stress  Nonadherence or noncompliance with instructions and pretest preparation  Undisclosed drug or alcohol use
  • 96. avoid costly mistakes  Communication errors account for more incorrect results than do technical errors.  Properly identify and label every specimen as soon as it is obtained.
  • 97. Educate the patient and family  Educate regarding the testing process and what will be expected  Record the date, time, type of teaching, information given, and person to whom the information was given.
  • 98. Educate the patient and family  Giving sensory and objective information that relates to what the patient will likely physically feel and the equipment that will be used is important so that patients can envision a realistic representation of what will occur.
  • 99. Educate the patient and family  Avoid technical and medical jargon  and adapt information to the patient's level of understanding.  Slang terms may be necessary to get a point across.
  • 100. Educate the patient and family  Encourage questions and verbalization of feelings, fears, and concerns  Do not dismiss, minimize, or invalidate the patient's anxiety  Develop listening skills, and be aware of nonverbal signals (ie, body language)
  • 101. Educate the patient and family  Above all, be nonjudgmental.  Emphasize that there is usually a waiting period (ie, turn-around time) before test results are relayed back to the clinicians and nursing unit.  Offer listening, presence, and support during this time of great concern and anxiety
  • 102. Educate the patient and family  Because of factors such as anxiety, language barriers, and physical or emotional impairments, the patient may not fully understand and assimilate instructions and explanations
  • 103. Educate the patient and family  To validate the patient's understanding of what is presented, ask the patient to repeat instructions given to evaluate assimilation and understanding of presented information.
  • 104. normal or reference values  Normal values are those that fall within 2 standard deviations (ie, random variation) of the mean value for the normal population.  Normal ranges can vary to some degree from laboratory to laboratory. Frequently, this is because of the particular type of equipment used
  • 105. normal or reference values  The reported reference range for a test can vary according to the laboratory used, the method employed, the population tested, and methods of specimen collection and preservation.
  • 106. normal or reference values  Interpretation of laboratory results must always be in the context of the patient's state of being.  Circumstances such as hydration, nutrition, fasting state, mental status, or compliance with test protocols are only a few of the situations that can influence test outcomes.
  • 107. clinical laboratory data values  may be reported in conventional units, SI units(Systéme International (SI) units), or both  The SI system uses seven dimensionally independent units of measurement to provide logical and consistent measurements
  • 108. clinical laboratory data values SI concentrations are written as amount per volume  (moles or millimoles per liter)  rather than as mass per volume (grams, milligrams, or milliequivalents per deciliter, 100 milliliters, or liter)
  • 109.  Numerical values may differ between systems or may be the same.  For example, chloride is the same in both systems: 95 to 105 mEq/L (conventional)  and 95 to 105 mmol/L (SI).
  • 110. Recognize margins of error  possibility exists that some tests will be abnormal owing purely to chance  because a significant margin of error arises from the arbitrary setting of limits.  Moreover, if a laboratory test is considered normal up to the 95th percentile, then 5 times out of 100, the test will show an abnormality even though a patient is not ill
  • 111. Cultural Sensitivity  Many cultures have diverse beliefs about diagnostic testing that requires blood sampling  Preserving the cultural well-being of any individual or group promotes compliance with testing and easier recovery from routine as well as more invasive and complex procedures
  • 112. END