Lyamuya E, Olausson-Hansson E, Albert J, Mhalu F, Biberfeld G
Department of Microbiology/Immunology, Muhimbili University College of Health Sciences, University of Dar es Salaam, Box 65001, Dar es Salaam, Tanzania.
J Clin Virol. 2000 Jun;17(1):57-63. doi: 10.1016/s1386-6532(00)00073-1.
In previous evaluations, the standard Amplicor HIV-1 DNA PCR test (Roche Diagnostic Systems) has been reported to have low sensitivity for the detection of some non-B HIV-1 subtypes. It has therefore become necessary to determine the performance of commercially available as well as prototype HIV-1 PCR assays for HIV-1 DNA detection in samples from various geographical settings, in order to assess their ability to detect the different HIV-1 genotypes.
To determine the performance of the prototype Roche Amplicor version 1.5 PCR test in comparison to that of the standard Roche Amplicor PCR test for the detection of HIV-1 DNA in blood samples from HIV-1 seropositive pregnant Tanzanian women infected with various HIV-1 subtypes.
This was a cross-sectional study done on 161 blood samples collected from 106 HIV-1 seropositive and 55 seronegative asymptomatic pregnant women attending antenatal clinic in Dar es Salaam, Tanzania.
Cell pellets for PCR were prepared from EDTA blood by the Amplicor whole blood PCR sample preparation method. Plasma was used for HIV serology by enzyme linked immunosorbent assays. Subtyping was done by the heteroduplex mobility assay (HMA) using cell pellets and/or plasma.
The sensitivities of the prototype PCR and the standard assays were 99.1% (105/106) and 97% (99/102), respectively. All samples from 55 HIV-1 seronegative women were negative by both PCR assays. Among the 101 samples subtyped by HMA, 48 (47%) were subtype A, 30 (30%) subtype C, 20 (20%) subtype D and 3 (3%) were indeterminate. In the standard DNA PCR assay, a statistically significantly higher proportion of subtype A samples had a low level of reactivity as measured as optical density compared with the subtypes C and D samples while in the prototype assay all three subtypes showed a high level of reactivity.
The Amplicor version 1.5 DNA PCR test has a high sensitivity for the detection of HIV-1 DNA in blood samples from Tanzanian adults. Since performance of this assay does not appear to be influenced by differences in HIV-1 subtypes A, C and D, it has the potential for use in the detection of HIV-1 DNA in samples from geographic areas where these subtypes are prevalent.
在以往的评估中，据报道标准的安普瑞可HIV-1 DNA聚合酶链反应检测（罗氏诊断系统）对某些非B型HIV-1亚型的检测灵敏度较低。因此，有必要确定市售以及原型HIV-1聚合酶链反应检测法在来自不同地理区域样本中检测HIV-1 DNA的性能，以评估它们检测不同HIV-1基因型的能力。
将罗氏安普瑞可1.5版原型聚合酶链反应检测法与标准罗氏安普瑞可聚合酶链反应检测法在检测感染各种HIV-1亚型的HIV-1血清学阳性坦桑尼亚孕妇血样中HIV-1 DNA的性能进行比较。
这是一项横断面研究，对从坦桑尼亚达累斯萨拉姆产前诊所就诊的106名HIV-1血清学阳性和55名血清学阴性无症状孕妇采集的161份血样进行研究。
采用安普瑞可全血聚合酶链反应样本制备方法从乙二胺四乙酸抗凝血中制备用于聚合酶链反应的细胞沉淀。采用酶联免疫吸附测定法检测血浆中的HIV血清学。使用细胞沉淀和/或血浆通过异源双链迁移率测定法（HMA）进行亚型分型。
原型聚合酶链反应检测法和标准检测法的灵敏度分别为99.1%（105/106）和97%（99/102）。55名HIV-1血清学阴性女性的所有样本两种聚合酶链反应检测法均为阴性。在通过HMA进行亚型分型的101份样本中，48份（47%）为A亚型，30份（30%）为C亚型，20份（20%）为D亚型，3份（3%）结果不确定。在标准DNA聚合酶链反应检测中，与C亚型和D亚型样本相比，A亚型样本中以光密度衡量的反应性水平在统计学上显著较低，而在原型检测中，所有三种亚型均显示出高反应性水平。
安普瑞可1.5版DNA聚合酶链反应检测法在检测坦桑尼亚成年人血样中HIV-1 DNA方面具有高灵敏度。由于该检测法的性能似乎不受HIV-1 A、C和D亚型差异的影响，它有潜力用于检测这些亚型流行的地理区域样本中的HIV-1 DNA。
