HIV Rapid Testing 1

Session 18
PREPARED BY TYSON JOEL SAMWEL
Learning Objectives
By the end of this session, students are expected to be able to:
• Define HIV rapid tests
• Describe HIV rapid tests currently used in Tanzania
• Describe the National HIV testing Algorithm
• Describe collection of blood for HIV Rapid Test
• Describe HIV testing using SD Bioline, Determine and Unigold
• Describe process of quality control in HIV testing
Definition of Term
• HIV rapid test: An immunochromatographic technique which is
used to detect HIV antibodies in blood.
• Specimens for this technique include whole blood, serum and
plasma.
• When performing HIV rapid test three blood samples must be
available; known HIV positive sample, known HIV negative
sample and a sample of unknown HIV status.
HIV Rapid Tests Currently Used in
Tanzania
• SD Bioline HIV-1/2
• Determine HIV-1/2
• Unigold HIV
HIV Rapid Tests are Divided into Two
Major Groups
• Antibody methods (to detect HIV)
• HIV Rapid tests
• Enzyme Linked Immunosorbent Assay (ELISA)
• Western Blot (confirmatory test if available)
• DNA PCR tests
• Useful in infant (<18 months) diagnosis
• The national protocol for PCR outlines the details of PCR
testing
• PCR can only be done at well-equipped laboratories at referral
hospitals
Principles of HIV Serological Tests
Principle of the test:
• An Immuno-Chromatographic Test (ICT) is used for the qualitative
detection of antibodies to HIV-1 and HIV-2 in blood. When a blood
sample is added to the sample pad it migrates through the
conjugate (colouring compound) and reconstitutes and mixes with
the conjugate. This mixture continues to migrate through the solid
phase to the immobilised antigens at the sample window site.
• If antibodies to HIV-1 and/or HIV-2 are present in the sample, they
bind to the conjugate and to the antigen at the client window site,
forming a red band. If antibodies to HIV-1 and/or HIV-2 are absent,
no red band is formed at the client window site.
Principles of HIV Serological Tests
cont..,,
• To ensure assay validity, a procedural control bar is
incorporated in the assay device.
• The test is based on antibody and antigen reaction,
interpretation is based on manufacturers manual
• Antibody + Antigen = Agglutination which is detected as a
colour change on the respective test (Reactive/ Positive
result)
• No Antibody + Antigen = No agglutination (no colours
change) (Non Reactive / Negative result)
Advantages of HIV Rapid Test
• Patients receive results in under 30 minutes
• Whole blood can be used (finger prick)
• Minimal skill and equipment required for accurate interpretation
• Tests are easy to use and read
• Most test kits can be stored at temperatures between 2-30 oC
• Small numbers of patients can be tested at once (unlike ELISA,
which requires a batch of samples)
Advantages of HIV Rapid Test cont…
• Are as accurate (sensitivity and specificity) and reliable as
ELISA:
• Sensitivity: The ability of a test to give a positive finding
when the tested person truly has the disease.
• Specificity: The ability of a test to give a negative finding
when the tested person is truly free of a disease. An ideal
test is one with a high specificity and high sensitivity.
• Rapid testing supports increased number of testing sites
Disadvantages of HIV Rapid Test
• They can’t determine Sero-status during window period
• On some occasions some individuals take longer to develop
antibodies. (During that time HIV rapid tests will be negative but
DNA/RNA PCR will demonstrate presence of the virus)
• There is a possibility that a rapid test will detect antibodies other
than those to HIV hence one positive test is not conclusive.
More tests are required for confirmation. Refer to sensitivity and
specificity.
Important Information before
Performing the Test
Before performing the test the tester should record/indicate/have the
following information;
• The test date
• Client identification
• Name of the person performing the test
• The name of the test
• The expiration date of the test kit
• Test lot and batch number of the test kit
• A standard operating procedure
• Results slip and register
The National HIV Testing Algorithm
National (Serial) Testing Algorithm
• An algorithm is a process, set of rules, or ‘step-by-step procedure for
reaching a clinical decision or diagnosis, often set out in the form of a
flow chart, in which the answer to each question determines the next
question to be asked’.
 Bioline Determine HIV Uni-Gold HIV HIV Status
HIV ½ 1/2
Non-reactive Negative

Reactive Reactive Positive

Reactive Non-reactive Non-reactive Negative

Reactive Non-reactive Reactive Positive

Collection of Blood for HIV Rapid Test
Materials and Supplies
• It is important to be prepared with all the supplies and materials
that are needed for HIV testing. These include:
• Gloves, Aprons or laboratory coats, Timer, clock, or wrist watch
with minute hand ,Transfer pipettes, pipette tips, HIV rapid testing
kits, Alcohol Cotton Gauze / wool, Sterile lancets and syringes
• Safety box/ sharps bin or disinfectant jar for lancets, Marker pen
for marking, labelling, Paper towels for bench coating, cleaning,
and hand washing, Antiseptic for hand washing
• Leak-proof bag for containing or moving labelled biohazard waste
for incineration, Disinfectant e.g. Jik, Clorox, Bandages or plasters
Protocol for Collecting Specimen for
HIV Rapid Test
• Finger-prick sample is recommended but if not tested in the
room, venous blood samples should be drawn carefully and
slowly and sent to the lab as quickly as possible.
• Stored in the refrigerator and NOT frozen.
• Sample tube should be labelled with, Patient identification
number, Collection date
Finger Prick
• Finger pricks should be done on the side of the finger, not in the
middle.
• The ring finger is the preferred finger for pricking because it has
a large surface area.
• Finger preparation for collecting blood through a finger prick
involves the following steps:
• Position hand palm-side up. Choose the ring finger, or
whichever finger is least calloused
• Apply intermittent pressure to the finger to help the blood to flow
Finger Prick cont…
• Clean the fingertip with alcohol. Start in the middle and work outward to
prevent contaminating the area. Allow the area to dry
• Hold the finger and firmly place a new sterile lancet off-centre on the
fingertip
• Firmly press the lancet to puncture the fingertip
• Wipe away the first drop of blood with a sterile gauze pad or cotton ball
• Collect the specimen. Blood may flow best if the finger is held lower
than the elbow
• Apply a gauze pad or cotton ball to the puncture site until the bleeding
stops
HIV Testing Using SD Bioline
SD Bioline HIV Rapid Test
Materials Required
• Bioline HIV1/2 KIT Standard operating procedure, Gloves Aprons or
laboratory coats, Timer, clock, or wrist watch with minute hand,
Alcohol, Cotton Gauze / wool Sterile lancets and syringes
• Safety box/ sharps bin or disinfectant jar for lancets Marker pen for
marking or labelling, Paper towels for bench coating, cleaning, and
hand washing, Antiseptic for hand washing,
• Leak-proof bag for containing or moving labelled biohazard waste for
incineration o Disinfectant e.g. Jik, Clorox, Bandages or plasters,
Register book
Procedure (SD Bioline HIV Rapid Test)
• Serum, plasma or whole blood is used
• Pipette required amount of specimen and add to the device well
(10µl for serum or plasma,20µl for whole blood)
• Add 3 – 4 drops of essay diluent
• Wait for 10-20 minutes
• Read the results
HIV Testing Using Determine
Determine HIV Rapid Test
• Materials Required
• Determine HIV1/2 KIT, Standard operating procedure, Chase buffer,
Gloves, Aprons or laboratory coats, Timer, clock, or wrist watch with
minute hand, Alcohol, Cotton Gauze / wool, Sterile lancets and
syringes, Safety box/ sharps bin, disinfectant jar for lancets
• Marker pen for marking or labelling, Paper towels for bench coating,
cleaning, and hand washing, Antiseptic for hand washing, Leak-proof
bag for containing or moving labelled biohazard waste for
incineration, Disinfectant e.g. Jik, Clorox o Bandages or plasters,
Register book
Procedure
• Collect test items and other necessary lab supplies
• Use one strip per test and be sure to preserve the lot number
on the remaining packet of strips
• Label the test strip with client identification number
• Pull off the protective foil cover
• Collect 50μl of specimen using a precision pipette or one drop
using a plastic transfer pipette
Procedure cont…
• Apply the specimen to the absorbent pad on the strip
• For whole blood only add one drop of chase buffer to the
specimen pad
• In that case, be sure to have on hand the chase buffer because
it is not included in the kit
• Wait 15 minutes (no longer than 60 minutes) before reading the
results
• Read and record the results and other pertinent information on
the worksheet
IMPORTANT
• It is important to be sure not to touch the testing area when performing
the test. Also, when tearing off one test strip it is easier to bend the
unopened strip back and forth, and then tear the strip away from the
others.
• Once the test strip is opened, the testing area should not be touched. It
is important to make sure that the sample is moving along the test strip
– this will take a moment.
• If not, add one more drop of buffer to the strip. Also, if a timer is not
available, a watch or clock on the wall can be used to monitor the time.
• It is a good practice to write down the starting time or projected end
time to help you keep track of time.
HIV Testing Using Unigold
Uni-Gold HIV Test
• Materials required
• Uni-Gold HIV1/2 KIT, Standard operating procedure, Gloves,
Aprons or laboratory coats, Timer, clock, or wrist watch with minute
hand, Alcohol, Cotton Gauze / wool, Sterile lancets and syringes,
Safety box/ sharps bin or disinfectant jar for lancets
• Marker pen for marking or labelling, Paper towels for bench
coating, cleaning, and hand washing, Antiseptic for hand washing,
Leak-proof bag for containing or moving labelled biohazard waste
for incineration, Disinfectant e.g. Jik, Clorox, Bandages or plasters,
Register book
Procedure
• Collect test items and other necessary laboratory supplies
• Remove device from package and label device with client identification
number
• Collect specimen using the disposable pipette
• Add two drops (approx. 60μl) of specimen to the sample port in the
device
• Add two drops (approx. 60μl) of the appropriate wash reagent to
sample port
• Wait for 10 minutes (no longer than 20 min.) before reading the results
• Read and record the results and other pertinent information on the
worksheet
Process of Quality Control in HIV
Testing
• Quality control (QC): Comprises measures for verifying that a
test is working. The measures are taken in order to monitor the
validity of the technical aspects of the test procedure. QC
includes the testing of samples with known results so as to
verify that the testing procedure and materials are working
properly.
Quality assessment
• Internal
• Onsite audit
• On-site audit is necessary in order to confirm that all
SOPs are adhered to, including QC, record-keeping and
observation of staff performance.
• A programme of on-site auditing should include a
standard checklist of testing service indicators.
• Proficiency testing
• Proficiency testing (PT) or EQA schemes involve the
distribution of small panels of samples with known results
to all testing sites.
Quality assessment cont….
• External
• Blinded rechecking
• Accomplished by forwarding 5%-10% positive and 5%
−10% of negative specimens for retesting at a higher level
laboratory (reference laboratory)
Key Points
• Effective HIV testing requires specific materials, supplies, and
preparation.
• HIV rapid test currently used in Tanzania are:
• Bioline, Determine, Uni-Gold
• HIV rapid testing should follow the National Algorithm.
• When performing HIV testing, adhere to standard operating
procedure.
• Recording of results is done in appropriate register books and
forms.
Evaluation
• What is National HIV testing Algorithm?
• What is a Standard Operating Procedure?
• Describe the collection of blood for HIV Rapid Test.
References
• CDC (2009). DPDx, Laboratory Identification of Parasites of Public Health
Concern http://www.dpd.cdc.gov/dpdx/HTML/Image_Library.htm/
• NACP, (2007) .HIV Rapid Test Training Package. Dar es Salaam, Tanzania:
National AIDS Control Programme, Ministry of Health & Social Welfare
• NACP/MOHSW. (2005) National Guidelines for Clinical Management of
HIV/AIDS. (2nd ed.) . Dar es Salaam, Tanzania: National AIDS Control
Programme, Ministry of Health & Social Welfare.
• NACP/MOHSW. (2005). National Guidelines for Voluntary Counselling & Testing.
Dar es Salaam, Tanzania: National AIDS Control Programme, Ministry of Health
& Social Welfare.
• NACP/NTLP. (2007). Manuals for In-service Training on Collaborative TB and HIV
activities. Dar es Salaam, Tanzania: National AIDS Control Programme and
National TB & Leprosy Control Programme, Ministry of Health & Social Welfare