|Year : 2013 | Volume
| Issue : 4 | Page : 417-418
Comparison of results obtained with different volumes of whole blood in human immunodeficiency virus-1 deoxyribonucleic acid polymerase chain reaction technique
D Anitha, SM Jacob
Department of Experimental Medicine, The Tamil Nadu Dr. MG Ramachandran Medical University, Chennai, Tamil Nadu, India
|Date of Submission||22-May-2013|
|Date of Acceptance||23-Aug-2013|
|Date of Web Publication||25-Sep-2013|
S M Jacob
Department of Experimental Medicine, The Tamil Nadu Dr. MG Ramachandran Medical University, Chennai, Tamil Nadu
Source of Support: None, Conflict of Interest: None
|How to cite this article:|
Anitha D, Jacob S M. Comparison of results obtained with different volumes of whole blood in human immunodeficiency virus-1 deoxyribonucleic acid polymerase chain reaction technique. Indian J Med Microbiol 2013;31:417-8
|How to cite this URL:|
Anitha D, Jacob S M. Comparison of results obtained with different volumes of whole blood in human immunodeficiency virus-1 deoxyribonucleic acid polymerase chain reaction technique. Indian J Med Microbiol [serial online] 2013 [cited 2020 May 30];31:417-8. Available from: http://www.ijmm.org/text.asp?2013/31/4/417/118885
Human Immunodeficiency virus-1 deoxyribonucleic acid (HIV-1 DNA) polymerase chain reaction (PCR) has been used in diagnosing HIV-1 in infants less than 18 months of age.  Roche Amplicor HIV-1 DNA Test version 1.5 is the most extensively used commercial kit worldwide.  Based on the kit insert, 500 μl of whole blood is required to perform the test. It is often difficult to obtain large volume of blood from infants. We had standardised and used in-house qualitative HIV-1 DNA PCR assay using 200 μl of whole blood and this has been previously published in this journal in 2008.  The objective of this study was to compare the results of different blood volumes-50 μl, 100 μl of whole blood as against the conventional 200 μl using in- house qualitative HIV-1 DNA PCR.
In this study, 85 infant samples were sent to our laboratory for PCR testing from various hospitals in South India and Mumbai. Qualitative in-house HIV-1 DNA PCR using 200 μl of whole blood was performed. DNA extraction was done using Qiagen Kit and PCR was performed with gag primers. The amplified products were run on gel electrophoresis and the positive bands were detected at 650 base pairs. Of the 85 samples, 55 HIV-1 positive and 30 HIV-1 negative samples were retested with 50 μl and 100 μl of whole blood.
The 85 infants' age ranged between 2 weeks and 16 months. Of the HIV positive infants, 30 were males and 25 females and of the HIV negative infants, 15 were males and 15 females. The concordance rate was 100% for 55 HIV-1 DNA PCR positive samples and 100% for 30 HIV-1 DNA PCR negative samples using 50 μl and 100 μl as against 200 μl [Table 1].
|Table 1: HIV-1 DNA PCR test results using different volume of whole blood|
Click here to view
HIV-1 DNA PCR is a very sensitive test and is able to detect 1-10 copies of proviral DNA. Our study revealed that there was no significant difference in the results when using 50 μl, 100 μl and 200 μl of whole blood for DNA extraction. DNA PCR results obtained using whole blood is the gold standard.  In a recent study from Tanzania, it was showed that the sensitivity of HIV-1 DNA PCR using 500 μl and 200 μl of venous blood was 95% and 98.3%, respectively, and specificity was 99.6% using both volumes.  Piwowar-Manning et al., obtained concordant results with 100 μl and 500 μl of whole blood using Roche Amplicor HIV-1 DNA PCR Assay version 1.5.  Therefore, lesser volumes of blood is sufficient to perform HIV-1 DNA PCR test and overcome difficulties encountered while performing venipuncture.
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