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Implementation of a Classic Nested PCR DNA for HIV Diagnosis in Kinshasa

Background: The interest in the technique of HIV diagnosis by Polymerase Chain Reaction (PCR) is in: (i) primary infection by the Human Immunodeficiency Virus (HIV), where it is necessary to identify the patient in time, (ii) screening of newborns for HIV-positive mothers, and (iii) the accuracy and reliability of the technique.

Objective: This study aims to assess the feasibility and the performance of the Nested PCR of Deoxyribose Nucleic Acid (DNA) for HIV diagnosis in Kinshasa in order to improve the detection and management of HIV infected patients.

Methods: The present study is a cross-sectional study in collaboration with 3 centers in Kinshasa. Our sample consisted of 171 individuals who have made voluntary testing for HIV. 5.0ml of blood was collected in tubes with anticoagulant, centrifuged for separation into 3 phases. 500μl of buffy coat was collected in a pre-labeled tube. The DNA extraction was made from 200μl of buffy coat using the QIAamp DNA Mini Kit from QIAGEN ® for DNA extraction. Classical HLA PCR and Nested PCR gag and pol were performed to determine the proviral DNA. Nested PCR on env was performed in cases of discordance of the results of gag and pol. The revelation was made under UV after electrophoretic migration on 1% agarose gel. The data were collected in confidentiality in the forms developed and pre-tested for the study, entered on Windows Excel 2007 and analyzed using SPSS 17.0 for Windows. The statistical significance was set at p <0.050.

Results: For Monkole, 22 samples were positive, 18 negative and 6 indeterminate for HIV. For Organization for Disease Prevention and Health Promotion (OPPS), we received 55 samples positive for HIV who served as a control for the study. For Foundation "La Grâce", 30 samples were positive, 25 negative and 15 indeterminate for HIV. This gives a total of 107 samples positive, 43 negative and 21 indeterminate for HIV. After amplification by Nested PCR DNA, we have a total of 112 samples positive and 59 HIV-negative. Of the 21 indeterminate samples, 3 were positive and 18 negative by PCR. 2 samples were negative by RDT but positive by PCR.

Conclusion: The Rapid Diagnostic Test (RDT) results and those of PCR DNA Nestled allowed us to evaluate and validate the technic of diagnosis of HIV in Kinshasa by this new technic. This method provides a more reliable alternative for the diagnosis of HIV Kinshasa.

Author(s): Erick Kamangu Ntambwe , Christian Mayemba , Samclide Mbikayi , Adolphe Ndarabu , Richard Kalala Lunganza , Georges Mvumbi Lelo , Dolores Vaira

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