APPLIED MICROBIOLOGY

LABORATORY DIAGNOSIS OF AIDS

Definition

Acquired immunodeficiency syndrome (AIDS) is a disease characterized by the occurrence of a life

threatening opportunistic infection or tumours or both. It is caused by a human retrovirus known as
human immunodeficiency virus (HIV).

CDC, Atlanta, USA has given a definition of AIDS according to which any person who is positive for
HIV antibody and who has CD4 cell count of < 200/ cubic ml, irrespective of his clinical status, shall
be considered a case with AIDS.

Clinical Samples

Though HIV is present in many body fluids such as saliva, tears etc; the most important of all these
is blood. About 5 ml of blood is collected aseptically using sterile and disposable syringe and needle
and is safely transported to the laboratory as early as possible. If delay is inevitable, sample can be
refrigerated. Transportation should be done on wet ice.

The laboratory diagnosis is based on various tests which are summarized in Table 82.8. The
diagnosis is based on antigen or antibody detection or showing the immunological abnormalities
caused by HIV infection.

TESTS FOR DETECTION

Immunological abnormality Antibody by ELISA
or Agglutination

Opportunistic infections Antigen by ELISA or PCR

Carcinomas and sarcomas Virus by co-culture method

Tests Indicative of Immunodeficiency

• Lowered total leucocyte count

• Lymphopenia
• Ratio of CD4:CD8 reversed
• CD4 cells <200/mL3
• Cutaneous anergy
• Elevated IgG and IgA
• Impaired blastogenesis by T and B cells

Opportunistic Infections and Malignancies/ Diseases suggestive of AIDS are:

PARASITIC
. Cryptosporidium
. Pneumocystis carinii pneumonia
. Toxoplasmosis of Nervous System
. Strongyloides stercoralis outside GIT FUNGAL
· Visceral candidiasis
· CNS cryptococcosis
BACTERIAL
Disseminated infection due to
- M. avium
- M. kansasii

VIRAL
· Systemic CMV infection
. Systemic herpes simplex virus infection
. Progressive multifocal leucoencephalopathy

MALIGNANCIES
· Kaposi's sarcoma
. Primary intracerebral lymphoma

Detection of Antibody

ELISA tests of different types are being used for antibody detection. ELISA tests are used as
screening procedures for HIV infection. A positive ELISA for HIV antibody can mean any of the
followings:
a. Current infection with HIV
b. Past exposure to HIV
c. False positive due to cross reaction
An ELISA that is negative for HIV antibody indicates any of the followings:
a. Absence of HIV infections
b. Failure of the test to pick up true positive and
c. Very early infection when antibodies are yet to be formed

Western blot test is used to confirm all those samples which are positive with ELISA. This test
detects antibodies to various components of HIV and hence is more specific.

Detection of Antigen

HIV antigen can be detected in serum, plasma, CSF, etc. using indirect ELISA tests. Though it is not
a widely used method because of its poor sensitivity, yet it has numerous advantages:
a. Active infection can be detected
b. Early diagnosis can be established
c. Diagnosis in newborns and infants can be made in spite of the presence of passively transferred
antibodies.
d. HIV infection can be distinguished from other immunodeficiency disorders.

Isolation of Virus is used as a routine diagnostic aid. The main use is to find out the prevalent HIV
strain and preparation of antigen against locally prevalent strain. It is done by the co-cultivation
technique using peripheral lymphocytes.

MOLECULAR LEVEL DETECTION

Highly sensitive and specific tests such as dot blot hybridisation, polymerase chain reaction (PCR)
etc are now being used by well equipped laboratories. PCR has been employed for different uses in
AIDS

Biosafety in Laboratory

Utmost biosafety is mandatory in laboratory where material potentially infected with HIV is being
handled. Use of disposable syringes and needles, avoiding mouth pipetting and handling of
potentially infected material in biosafety cabinets is essential. The procedures by which HIV can be
inactivated include boiling, treating with 70% ethanol, 20% gluteraldehyde, 5% formaldehyde and
0.5% lysol solution.