Detection of HIV proviral DNA by a duplex fluorescence PCR for early diagnosis of HIV infection in infants

Objective To establish a duplex fluorescence PCR for detection of HIV proviral DNA and to evaluate its application for early diagnosis of HIV infection in infants. Methods A duplex fluores- cence PCR system was set up based on TaqMan technology for detection of human ribonuclease P （ RNase P） gene and long terminal repeat （LTR） region of HIV. A recombinant plasmid containing the targeted gene fragment, pTG19-T, was constructed by TA cloning technique and used as the template for evaluation of sen- sitivity of the assay. Blood samples from 11 healthy individuals and 98 HIV-infected patients were collected and detected to validate the assay specificity. The assay of duplex fluorescence PCR was then carried out to detect 96 infant blood samples collected from several maternal and child health hospitals in Zhejiang province from January 2011 to September 2012 for early diagnosis of HIV infection. The results were compared with those by using the Roche HIV DNA qualitative detection kit. Results The established duplex fluorescence PCR could specifically detect HIV proviral DNA with a specificity of 100% and a detection sensitivity of 100 cps per reaction. The coincidence rate between the established assay and the Roche HIV DNA qualitative de- tection kit was 100% in the detection of 96 blood samples. Conclusion The duplex fluorescence PCR as- say showed advantages of cost-effectiveness, convenience, good specificity and accuracy with high sensitivi- ty. It could be used for early diagnosis of HIV infection in infants and also as a general technical platform for the detection of HIV proviral DNA. Key words: Fluorescence PCR;  Human immunodeficiency virus （HIV） ;  Infant;  Early diagnosis