Abstract

 Background and Objectives

 Human Immunodeficiency Virus type 1 (HIV-1) is the causative agent of Acquired Immunodeficiency Syndrome (AIDS) in man and diagnosis of HIV-1 genome in suspicious specimens is of special importance. Viral transmission through blood and blood products during the window period is considered to be an important concern. In addition, the employment of rapid, sensitive and accurate techniques is highly necessary for diagnosis of HIV-1 prior to antibody production in infants born from infected mothers. In the present study, a sensitive and rapid RT-Nested PCR to detect viral genome has developed.

 Materials and Methods

 In this study, a sensitive RT-Nested PCR technique for detection of a conserved HIV-1 "gag gene" sequence was developed. By using a specific primer pair, the fragment was amplified in two rounds of PCR. In order to confirm positive results, the developed technique was applied to standard Iranian panel as well as to positive specimens from different infection and disease categories in which reactivity was proven by confirmatory HIV-1 tests (ELISA and western blot).

 Results

 Thirty five sera from different stages of the disease as well as 15 standard Iranian panels and 20 negative control sera were collected and tested by the developed technique. In positive cases, a specific band was observed on agarose gel electrophoresis, while no band could be detected for negative control sera.

 Conclusions

 In this study, it was demonstrated that the developed HIV-1 RT-Nested PCR has a high sensitivity and specificity for diagnosis of HIV-1 infection. It has the advantage of viral genome detection prior to seroconversion and can be easily applied to detect HIV-1 infection during the window period.

 �

 Key words : Nested PCR , RT-PCR, HIV-1, Gag gen�