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1.Laboratory diagnosis of HIV

 The diagnosis of HIV infection is established by one of the
following methods:
 detecting antibodies to the virus(ELISA, rapid test
&western blotting).
 detecting the viral p24 antigen
 detecting viral nucleic acid (southern or Northern
blootting).
 culturing HIV

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  Viral load
 Marker of HIV replication rate
 Number of HIV RNA copies/mm3 plasma
 CD4 count
 Marker of immunologic damage
 Number of CD4 T-lymphocytes cells/mm3 plasma
 Normal value:500 -1400 cells/mm3

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detection of antibody to HIV
 By far, the most widely used test is the detection of
antibody to HIV
 Antibodies to gp41 and p24 antigens are the first
detectable serologic markers following HIV infection.
 IgG antibodies appear 6 to 12 weeks following HIV
infection in the majority of patients and by six months
in 95 percent of patients.
 Positive tests should be confirmed with repeat tests or
corroborating laboratory data.

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 Common HIV Antibody Tests
A. Enzyme Linked Immunosorbent Assays (ELISA)

B. Rapid Tests

C. Western Blot

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Rapid Tests
 Rapid tests are useful for small laboratories that routinely

perform fewer than 100 HIV tests per day, for laboratories
without electricity or equipment, and for geographic areas with
limited laboratory infrastructure.

 In some instances, even if a laboratory performs more than 100

tests per day but only during a limited time in a year, rapid tests
may be more appropriate than ELISA.

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  A result can usually be obtained in less than 45 minutes, and
it is easy to interpret.
 Training is required to correctly perform the test and
interpret the results.
 The test kits generally contain all reagents needed to run the
assay, no additional reagents or equipment is required.
 Many rapid tests do not require electricity, special
equipment, refrigeration, or highly skilled staff although a
few require refrigeration for heat- sensitive reagents.

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  Sensitivity approaches 100%; specificity is >99%

 Negative tests can be reported as negatives

 Positive results should be confirmed

 Useful in situations where immediate results are important to

manage decisions

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 Previous rapid test algorithm of HIV
Blood
Blood Sample
Sample

Test 1 (wantiebejing)
(Determine)

Non
Non -reactive
-reactive Reactive
Reactive

Report
Report Negative
Negative Test 2 (unigold)
(Capillus)

Reactive
Reactive
Non
Non Reactive
Reactive
Report
Report Positive
Positive

Test
Test 33 (vikia)
(Unigold)
(Unigold)

Reactive
Reactive Result
Result Non
Non -reactive
-reactive Result
Result

Report
Report Positive
Positive Report
Report Negative
Negative
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Current National HIV Testing Algorithm (NHTA) in Ethiopia manual 2018. (Key:
A1 = Assay1, A2 = Assay 2, A3 = Assay 3. * Brand name of HIV rapid test kit
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 Serology of hepatitis tests

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serological diagnosis of HBV
1. Markers for hepatitis B virus
 Antigens
Hepatitis B surface antigen (HBsAg)
 Detected during incubation period
 Found during active phase of the disease
 It persists for months and years, the individual is carrier and
potentially infectious.
 Produced in the cytoplasm of infected hepatocytes
 It is the outer lipoprotein coat (envelope)

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 Serology of Hepatitis virus
Hepatitis B core antigen (HBcAg)
 It is the core of HBV
 It is located in the nuclei of
hepatocytes
 It is not detectable in serum
Hepatitis B e antigen (HBeAg)
 It is a minor component of the
viron
 It is found during acute
infection and then usually
disappear and usually reliable
diagnostic indication of active
infection.
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 Serology of HBV
 Antibodies
 Anti – HBsAg
 Appears after disappearance of HBsAg
 Shows past infection and immunity
 It is a measure of recovery from HBV infection
 In carriers, HBsAg and anti–HbsAg does not develop

 Anti-HBeAg
 Not frequently observed in patient with chronic infection

 Patients with anti HBeAg are usually not infectious because they have
low titer of HBV.

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 Serology of HBV
Anti -HBcAg

 Is actively infected individuals antibodies to HBcAg appear in

serum after the appearance of HBsAg before the onset of

symptoms.

 Reliable marker of recent infection

 Absences of IgM anti HBcAg in HBsAg positive individuals

indicates , it shows carrier state.

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 Serology of HBV
 A battery of serological tests are used for the diagnosis
of acute and chronic hepatitis B infection.
 HBsAg used as a general marker of infection.
 HBsAbused to document recovery and/or immunity to
HBV infection.
 anti-HBc IgMmarker of acute infection.
 anti-HBcIgGpast or chronic infection.
 HBeAgindicates active replication of virus and
therefore infectiveness.

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 Sero of HBV
 Anti-Hbevirus no longer replicating.
 However, the patient can still be positive for HBsAg
which is made by integrated HBV.
 HBV-DNAindicates active replication of virus,
more accurate than HBeAg especially in cases of
escape mutants.
 Used mainly for monitoring response to therapy.

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 Serological diagnosis of Hepatitis C
virus
Antibody testing for HCV
 The diagnosis of hepatitis C infection is usually made
serologically by detecting anti-HCV IgG in serum.
 Most patients infected with HCV become carriers.
 Antibody is detectable 6–8 weeks after infection.
 Detecting anti-HCV antibody is also used to screen donor
blood.
 ELISA and rapid anti-HCV antibody tests to diagnose
hepatitis C and screen blood for HCV are commercially
available.
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 Hepatitis C Virus Infection
Typical Serologic Course
anti-HCV

Symptoms

Titre

ALT

Normal

0 1 2 3 4 5 6 1 2 3 4
Months Years

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 Serology diagnosis of HCV
 HCV antibody - generally used to diagnose hepatitis C infection.
Not useful in the acute phase as it takes at least 4 weeks after
infection before antibody appears.
 HCV-RNA - various techniques are available e.g. PCR and
branched DNA. May be used to diagnose HCV infection in the
acute phase.
 However, its main use is in monitoring the response to antiviral
therapy.
 HCV-antigen - an EIA for HCV antigen is available.

 It is used in the same capacity as HCV-RNA tests but is much

easier to carry out.

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 Serology of HAV,HDV,HEV

HAV, HDV, HEV

 Hepatitis A can be diagnosed serologically by detecting HAV-

specific IgM antibody which appears in the serum at the onset

of jaundice and persists for about 10 weeks.

 Antigen and antibody tests are available to diagnose hepatitis D

and hepatitis E, but the assays are expensive and usually

performed only in specialist laboratories.

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 HBV - HDV Co-infection
Typical Serologic Course
Symptoms

ALT Elevated

Titre
anti-HBs
IgM anti-HDV

HDV RNA

HBsAg
Total anti-HDV

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Time after Exposure
 HBV - HDV Super-infection
Typical Serologic Course
Jaundice

Symptoms

Total anti-HDV
ALT
Titre

HDV RNA
HBsAg

IgM anti-HDV

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 Hepatitis E Virus Infection
Typical Serologic Course
Symptoms

ALT IgG anti-HEV

Titer IgM anti-HEV

Virus in stool

0 1 2 3 4 5 6 7 8 9 1 1 1 1
0 1 2 3
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Weeks after Exposure
 Reading assignment

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