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首页> 外文期刊>BMC Pediatrics >Evaluation of a single round polymerase chain reaction assay using dried blood spots for diagnosis of HIV-1 infection in infants in an African setting
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Evaluation of a single round polymerase chain reaction assay using dried blood spots for diagnosis of HIV-1 infection in infants in an African setting

机译：评价使用干血斑的单轮聚合酶链反应测定法在非洲环境中诊断婴儿的HIV-1感染

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Background The aim of this study was to develop an economical 'in-house' single round polymerase chain reaction (PCR) assay using filter paper-dried blood spots (FP-DBS) for early infant HIV-1 diagnosis and to evaluate its performance in an African setting. Methods An 'in-house' single round PCR assay that targets conserved regions in the HIV-1 polymerase (pol) gene was validated for use with FP-DBS; first we validated this assay using FP-DBS spiked with cell standards of known HIV-1 copy numbers. Next, we validated the assay by testing the archived FP-DBS (N = 115) from infants of known HIV-1 infection status. Subsequently this 'in-house' HIV-1 pol PCR FP-DBS assay was then established in Nairobi, Kenya for further evaluation on freshly collected FP-DBS (N = 186) from infants, and compared with findings from a reference laboratory using the Roche Amplicor? HIV-1 DNA Test, version 1.5 assay. Results The HIV-1 pol PCR FP-DBS assay could detect one HIV-1 proviral copy in 38.7% of tests, 2 copies in 46.9% of tests, 5 copies in 72.5% of tests and 10 copies in 98.1% of tests performed with spiked samples. Using the archived FP-DBS samples from infants of known infection status, this assay was 92.8% sensitive and 98.3% specific for HIV-1 infant diagnosis. Using 186 FP-DBS collected from infants recently defined as HIV-1 positive using the commercially available Roche Amplicor v1.5 assay, 178 FP-DBS tested positive by this 'in-house' single-round HIV-1 pol PCR FP-DBS PCR assay. Upon subsequent retesting, the 8 infant FP-DBS samples that were discordant were confirmed as HIV-1 negative by both assays using a second blood sample. Conclusions HIV-1 was detected with high sensitivity and specificity using both archived and more recently collected samples. This suggests that this 'in-house' HIV-1 pol FP-DBS PCR assay can provide an alternative cost-effective, reliable and rapid method for early detection of HIV-1 infection in infants.
机译：背景技术这项研究的目的是开发一种经济的“内部”单轮聚合酶链反应（PCR）检测方法，该方法使用滤纸干燥血斑（FP-DBS）进行早期婴儿HIV-1诊断，并评估其在儿童中的表现。非洲环境。方法验证了针对HIV-1聚合酶（pol）基因保守区域的“内部”单轮PCR检测可用于FP-DBS。首先，我们使用掺有已知HIV-1拷贝数的细胞标准物的FP-DBS验证了该测定方法。接下来，我们通过测试已知HIV-1感染状况婴儿的FP-DBS（N = 115）来验证该检测方法。随后，在肯尼亚内罗毕建立了这种“内部” HIV-1 pol PCR FP-DBS分析方法，以进一步评估从婴儿身上新鲜收集的FP-DBS（N = 186），并与使用该方法的参考实验室的结果进行比较。罗氏Amplicor ？ HIV-1 DNA测试，1.5版检测。结果HIV-1 pol PCR FP-DBS检测可以在38.7％的检测中检测到1个HIV-1前病毒拷贝，在46.9％的检测中检测到2个拷贝，在72.5％的检测中检测5个拷贝，在98.1％的检测中检测10个拷贝。加标样品。使用已知感染状态婴儿的存档FP-DBS样本，该测定对HIV-1婴儿诊断的敏感性为92.8％，特异性为98.3％。使用市售的Roche Amplicor v1.5测定法从最近定义为HIV-1阳性的婴儿中收集的186种FP-DBS，通过此“内部”单轮HIV-1 pol PCR FP-DBS测试了178种FP-DBS呈阳性PCR测定。在随后的重新测试中，使用第二份血液样本的两种测定均将不一致的8例婴儿FP-DBS样本确认为HIV-1阴性。结论使用已存档和最近收集的样本均以高灵敏度和特异性检测到HIV-1。这表明，这种“内部” HIV-1 pol FP-DBS PCR检测方法可以为早期检测婴儿的HIV-1感染提供另一种经济高效，可靠且快速的方法。

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