Dunne Amanda L, Mitchell Fiona, Allen Kelly M, Baker H W Gordon, Garland Suzanne, Clarke Gary N, Mijch Anne, Crowe Suzanne M
Macfarlane Burnet Institute for Medical Research and Public Health, PO Box 2284, Melbourne, VIC 3001, Australia.
J Clin Virol. 2003 Feb;26(2):239-45. doi: 10.1016/s1386-6532(02)00122-1.
With HIV-1-infected individuals now facing the prospect of relatively long and healthy lives, many discordant couples (where the male is HIV-1 seropositive) are seeking to have children. To assist reducing the risks of heterosexual and subsequent vertical transmission in this situation, quantification of HIV-1 viral load in seminal plasma may be effective as one of several measures to reduce the risk of infecting the mother during insemination, potentially providing a better indication of infectivity than blood plasma analysis.
OBJECTIVE(S): To modify existing molecular methods for the purpose of analysing HIV-1 viral load in seminal plasma.
Two commercial assays for HIV-1 RNA quantification were used to assess their sensitivity, specificity and precision for quantification of seminal plasma samples. Seminal plasma samples were prepared with an additional centrifugation step to aid removal of inhibitors to molecular assays.
Seminal plasma samples exhibited specificity of >95%, equivalent to that reported by the manufacturers of the commercial assays. With additional centrifugation, complete inhibition of 2/19 (10%) seminal plasma samples was observed using the RT-PCR assay, and inhibition was not apparent in the bDNA assay. Quantification of HIV-1 RNA in seminal plasma samples in both assays was equivalent to that observed in plasma samples and did not appear to be affected by the additional centrifugation step.
Minor modification of the RT-PCR assay procedure by additional centrifugation of seminal plasma improved the sensitivity of the assay. Inhibition was not apparent with the bDNA assay.
随着感染人类免疫缺陷病毒1型（HIV-1）的个体如今面临着相对长寿且健康的前景，许多配偶双方一方感染（男性为HIV-1血清反应阳性）的夫妇都希望生育子女。为了帮助降低这种情况下异性传播及后续垂直传播的风险，对精液血浆中的HIV-1病毒载量进行定量分析可能是有效的措施之一，相比血浆分析，它可能更能准确指示传染性，从而降低授精过程中感染母亲的风险。
改进现有的分子方法以分析精液血浆中的HIV-1病毒载量。
使用两种用于HIV-1 RNA定量分析的商业检测方法，评估它们对精液血浆样本定量分析的灵敏度、特异性和精密度。通过额外的离心步骤制备精液血浆样本，以帮助去除对分子检测有抑制作用的物质。
精液血浆样本的特异性>95%，与商业检测方法制造商报告的结果相当。经过额外离心后，使用逆转录聚合酶链反应（RT-PCR）检测方法时，观察到19份精液血浆样本中有2份（10%）完全受到抑制，而在分支DNA（bDNA）检测方法中未观察到抑制现象。两种检测方法对精液血浆样本中HIV-1 RNA的定量分析结果与血浆样本中的结果相当，且似乎不受额外离心步骤的影响。
通过对精液血浆进行额外离心对RT-PCR检测程序进行微小改进，提高了该检测方法的灵敏度。bDNA检测方法未出现明显抑制现象。
