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CORRESPONDENCE
Year : 2013  |  Volume : 31  |  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


Department of Experimental Medicine, The Tamil Nadu Dr. MG Ramachandran Medical University, Chennai, Tamil Nadu, India

Date of Submission22-May-2013
Date of Acceptance23-Aug-2013
Date of Web Publication25-Sep-2013

Correspondence Address:
S M Jacob
Department of Experimental Medicine, The Tamil Nadu Dr. MG Ramachandran Medical University, Chennai, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0255-0857.118885

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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 2019 Jun 24];31:417-8. Available from: http://www.ijmm.org/text.asp?2013/31/4/417/118885


Dear Editor,

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. [1] Roche Amplicor HIV-1 DNA Test version 1.5 is the most extensively used commercial kit worldwide. [2] 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. [3] 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

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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. [4] 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. [5] 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. [6] Therefore, lesser volumes of blood is sufficient to perform HIV-1 DNA PCR test and overcome difficulties encountered while performing venipuncture.

 
 ~ References Top

1.Sherman GG, Stevens G, Jones SA, Horsfield P, Stevens WS. Dried blood spots improve access to HIV diagnosis and care for infants in low-resource settings. J Acquir Immune Defic Syndr 2005;38:615-7.  Back to cited text no. 1
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2.Stevens W, Sherman G, Downing R, Parsons LM, Ou CY, Crowley S, et al. Role of the laboratory in ensuring global access to ARV treatment for HIV-infected children: consensus statement on the performance of laboratory assays for early infant diagnosis. Open AIDS J 2008;2:17-25.  Back to cited text no. 2
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3.Jacob SM, Anitha D, Viswanath R, Parameshwari S, Samuel NM. The use of dried blood spots on filter paper for the diagnosis of HIV-1 in infants born to HIV seropositive women. Indian J Med Microbiol 2008;26:71-4.  Back to cited text no. 3
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4.Susanna C, Fulvio E, Davide B, Richard L, Mamy AP, Giovanna P, et al. Evaluation of different deoxyribonucleic acid (DNA) extraction methods using dried blood spot for early infant diagnosis of HIV1 in Sub-Saharan Africa. Afr J Microbiol Res 2012;6:5609-17.  Back to cited text no. 4
    
5.Nsojo A, Aboud S, Lyamuya E. Comparative evaluation of amplicor HIV-1 DNA test, version 1.5, by manual and automated DNA extraction methods using venous blood and dried blood spots for HIV-1 DNA PCR testing. Tanzan J Health Res 2010;12:229-35.  Back to cited text no. 5
    
6.Piwowar-Manning E, Lugalia L, Kafufu B, Jackson JB. Comparison of results obtained with amplicor HIV-1 DNA PCR test version 1.5 using 100 versus 500 microliters of whole blood. J Clin Microbiol 2008;46:1104-5.  Back to cited text no. 6
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