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目的建立鼠艾滋病模型血浆病毒载量荧光定量聚合酶链(PCR)检测方法。方法将体外合成的鼠艾滋病病毒(FLV)核糖核酸(RNA)标准品稀释为10个浓度梯度,进行逆转录,PCR荧光定量检测,经批内与批间差异分析,再以PCR荧光定量检测不同倍数FLV脾悬液FLV拷贝数及相应被注射小鼠模型的血浆病毒载量,计算脾指数。结果研究设计的荧光定量PCR方法能检测到20～2×108copiesFLVRNA逆转录反应体系,在5个注射的FLV拷贝数范围内,血浆病毒载量与脾指数均不与初始注射的FLV拷贝数成正比,二者亦不呈现平行关系。结论研究采用的FLV荧光定量PCR方法敏感性好,特异性强,重现性好,显著提高FLV病毒的检测水平。
Objective To establish a real-time fluorescent quantitative polymerase chain reaction (PCR) method for the determination of plasma viral load in murine models of AIDS. Methods FLV RNA standard was diluted to 10 concentration gradients in vitro and then was reverse transcribed and detected by PCR fluorescence quantitative analysis. The intra-and inter-assay differential analysis was used to detect FLV RNA. Fold FLV FLV copy number of spleen suspension and the corresponding injected mouse model of plasma viral load, calculate the spleen index. Results The fluorescence quantitative PCR method was able to detect 20 ~ 2 × 108 copies of FLV RNA reverse transcription reaction system. In the 5 injected FLV copy number range, neither the plasma viral load nor the spleen index was directly proportional to the initial injected FLV copy number , The two do not show a parallel relationship. Conclusion The FLV fluorescence quantitative PCR method used in this study is sensitive, specific and reproducible, and significantly improves the detection level of FLV virus.