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Early diagnosis of HIV-1: infected infants in Brazil using nested-PCR
R M Molina1, A D C Toro, M T N Silva
1Universidade Estadual de Campinas, Unicamp, São Paulo, Brazil.
Insights
Polymerase chain reaction (PCR) is a key test for early human immunodeficiency virus type 1 (HIV-1) diagnosis in infants. This study standardized PCR methods, confirming its effectiveness for detecting HIV-1 in infants suspected of perinatal infection.
Area of Science:
- Medical Virology
- Molecular Diagnostics
- Pediatric Infectious Diseases
Background:
- Early diagnosis of human immunodeficiency virus type 1 (HIV-1) in infants is crucial for timely intervention.
- Perinatal transmission remains a significant route of HIV-1 infection in newborns.
- Polymerase chain reaction (PCR) has emerged as a highly sensitive method for infant HIV-1 diagnosis.
Purpose of the Study:
- To standardize and evaluate the performance of a two-step Polymerase Chain Reaction (PCR) assay for the early diagnosis of HIV-1 infection in infants.
- To assess the sensitivity of different primer sets and target regions (gag, env, pol) for HIV-1 detection in pediatric samples.
Main Methods:
- Standardization of a two-step PCR amplification protocol for HIV-1 detection.
- Analysis of 41 infants under 18 months old suspected of perinatal HIV-1 infection.
- Utilized specific outer and inner primer sets (JA4-JA7, JA9-JA12, JA13-JA16, JA17-JA20 and JA5-JA6, JA10-JA11, JA14-JA15, JA18-JA19) targeting HIV-1 DNA.
Main Results:
- The two-step PCR assay was successfully standardized for HIV-1 detection in infants.
- Out of 41 infants analyzed, 12 tested positive for HIV-1 infection.
- The gag, env (region 1), and pol gene regions demonstrated higher sensitivity for HIV-1 detection.
Conclusions:
- The standardized two-step PCR method is effective for the early diagnosis of HIV-1 in infants.
- PCR is confirmed as the recommended diagnostic tool for identifying HIV-1 perinatal infections in infants.
- The study highlights the utility of specific gene regions for enhanced diagnostic sensitivity.
Abstract:
The polymerase chain reaction (PCR) has been the most promising test for HIV-1 early diagnosis in infants suspected of perinatal transmission. The first and second reactions of the amplification in 41 infants (under 18 months old) suspected of HIV-1 perinatal infection, were standardized and carried out in the present study. The first and the second PCR were carried out with the sets of primers JA4-JA7, JA9-JA12, JA13-JA16, and JA17-JA20 for the first reaction of amplification (outer primers) and JA5-JA6, JA10-JA11, JA14-JA15, and JA18-JA19 for the second reaction of amplification (inner primers), resulting in amplification of 131, 341, 172, and 129 pb, respectively. From 41 patients analysed, 12 patients presented positive to HIV-1 infection by PCR. The gag, env (region 1), and pol regions presented a greater sensitivity. The first and the second reactions of the amplification were performed with the same concentration of MgCl2 for all sets of primers. The results agree with several studies that affirm that the PCR is the indicated method for HIV-1 early diagnosis in infants suspected of perinatal infection.
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