Jacob S Mini, Anitha D, Vishwanath R, Parameshwari S, Samuel N M
Department of Experimental Medicine, The Tamilnadu Dr. MGR Medical University, Guindy, Chennai-32, Tamil Nadu, India.
Indian J Med Microbiol. 2008 Jan-Mar;26(1):71-4. doi: 10.4103/0255-0857.38864.
Polymerase chain reaction (PCR) is the most sensitive test to diagnose HIV-1 infection among infants born to HIV seropositive mothers. The purpose of this study was to evaluate the use of dried blood spot (DBS) specimens for PCR and to compare it with whole-blood stored in tubes for HIV-1 DNA PCR. Five hundred and seventy-seven whole-blood infant samples were tested using HIV-1 qualitative in-house nested DNA PCR. Three hundred and fifty-nine samples were from infants at 48 hours of birth and 218 samples at second month. All positive samples tested from whole-blood and every fifth negative sample were coated onto filter paper. DNA was extracted from the filter paper and was amplified using in-house nested PCR. Among the whole-blood samples tested using HIV-1 DNA PCR, 19 of 359 (5.29%) samples were HIV-1 positive and 340 (94.7%) were negative at 48 hours of birth. At second month, 19 (8.7%) of the 218 samples were positive and 199 (91.2%) were negative. Using dried filter paper, 18 samples (95%) tested positive from 19 positive samples (using whole-blood) and 1 tested negative at 48 hours of birth. The 68 negative samples tested using whole-blood were also negative in the DBS test (sensitivity 95% and specificity 100%). At second month, 19 were positive and 40 samples (every fifth sample of 199) were negative (sensitivity and specificity, 100%). PCR performed using DNA extracted from filter paper permits the diagnosis of HIV-1 infection among infants born to HIV-1 seropositive mothers. This assay is simple, rapid, sensitive and specific and can be used in resource limited settings.
聚合酶链反应(PCR)是诊断HIV血清阳性母亲所生婴儿感染HIV-1的最敏感检测方法。本研究的目的是评估干血斑(DBS)标本用于PCR的情况,并将其与用于HIV-1 DNA PCR的试管储存全血进行比较。使用HIV-1定性内部嵌套DNA PCR检测了577份婴儿全血样本。359份样本来自出生48小时的婴儿,218份样本来自第二个月的婴儿。所有全血检测出的阳性样本以及每五个阴性样本中的一个被涂覆在滤纸上。从滤纸上提取DNA,并使用内部嵌套PCR进行扩增。在使用HIV-1 DNA PCR检测的全血样本中,出生48小时时,359份样本中有19份(5.29%)HIV-1呈阳性,340份(94.7%)呈阴性。在第二个月,218份样本中有19份(8.7%)呈阳性,199份(91.2%)呈阴性。使用干滤纸,19份全血阳性样本中有18份(95%)检测呈阳性,出生48小时时1份检测呈阴性。使用全血检测的68份阴性样本在DBS检测中也呈阴性(敏感性95%,特异性100%)。在第二个月,19份呈阳性,40份样本(199份样本中的每五个样本)呈阴性(敏感性和特异性均为100%)。使用从滤纸上提取的DNA进行PCR可诊断HIV血清阳性母亲所生婴儿的HIV-1感染。该检测方法简单、快速、灵敏且特异,可用于资源有限的环境。