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用于检测坦桑尼亚成年人血液样本中1型人类免疫缺陷病毒DNA的内部和商业样本制备及PCR扩增系统的比较

Comparison of in-house and commercial sample preparation and PCR amplification systems for detection of human immunodeficiency virus type 1 DNA in blood samples from Tanzanian adults.

作者信息

Lyamuya E, Bredberg-Rådén U, Albert J, Grankvist O, Msangi V, Kagoma C, Mhalu F, Biberfeld G

机构信息

Department of Microbiology/Immunology, Muhimbili University College of Health Sciences, University of Dar es Salaam, Tanzania.

出版信息

J Clin Microbiol. 1997 Jan;35(1):278-80. doi: 10.1128/jcm.35.1.278-280.1997.

Abstract

This study compared the performance of several in-house nested PCR systems and the Amplicor human immunodeficiency virus type 1 (HIV-1) PCR kit in the detection of HIV-1 DNA in Tanzanian samples prepared by two different methods. All six of the in-house primer sets evaluated had a higher sensitivity for HIV DNA detection in samples prepared by the Amplicor PCR sample preparation method than in those prepared by the Ficoll-Isopaque (FIP) density gradient centrifugation method. A sensitivity of 100% was achieved by combining two in-house primer sets. The sensitivity of the standard Amplicor HIV-1 PCR kit was only 59%, whereas a modified Amplicor HIV-1 PCR test had a sensitivity of 98%. Our data show that Tanzanian samples prepared by the Amplicor preparation method are more suitable for HIV-1 PCR testing than samples prepared by the FIP method. The modified, but not the standard, Amplicor HIV-1 PCR kit provides an alternative to the nested in-house PCR technique for the diagnosis of HIV infection.

摘要

本研究比较了几种内部巢式PCR系统和Amplicor 1型人类免疫缺陷病毒(HIV-1)PCR试剂盒在检测通过两种不同方法制备的坦桑尼亚样本中HIV-1 DNA的性能。所评估的所有六种内部引物组对通过Amplicor PCR样本制备方法制备的样本中HIV DNA检测的灵敏度高于通过菲可-异泛影葡胺(FIP)密度梯度离心法制备的样本。通过组合两种内部引物组实现了100%的灵敏度。标准的Amplicor HIV-1 PCR试剂盒的灵敏度仅为59%,而改良的Amplicor HIV-1 PCR检测的灵敏度为98%。我们的数据表明,通过Amplicor制备方法制备的坦桑尼亚样本比通过FIP方法制备的样本更适合用于HIV-1 PCR检测。改良而非标准的Amplicor HIV-1 PCR试剂盒为HIV感染的诊断提供了一种替代巢式内部PCR技术的方法。

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