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目的:建立重组猿免疫缺损病毒-人色素上皮衍生因子(simian immunodeficiency virus-human pigment epithelium-derived factor,SIV-hPEDF)注射液中复制型SIV的实时定量PCR检测方法。方法:在提取重组SIV-hPEDF注射液供试品基因组RNA后,采用实时定量PCR的方法,检测供试品中的复制型SIV。结果:经过3次实验,所得TAT RNA标准品标准曲线均线性良好,线性方程为Y=-3.248lgX+38.12,相关系数为0.994;重组SIV-hPEDF注射液供试品中复制型SIV检测结果均为阴性。结论:所建立的复制型SIV实时荧光定量PCR检测方法重复性好、灵敏、准确,可用于重组SIV-hPEDF注射液中复制型SIV的检测。
OBJECTIVE: To develop a real-time quantitative PCR assay for replicating SIV in simian immunodeficiency virus-human pigment epithelium-derived factor (SIV-hPEDF) injection. Methods: After extracting the recombinant SIV-hPEDF injection for the test sample genomic RNA, real-time quantitative PCR was used to detect the replicative SIV in the test sample. Results: After three experiments, the standard curve of TAT RNA standard was linear, the linear equation was Y = -3.248lgX + 38.12, the correlation coefficient was 0.994. The results of recombinant SIV-hPEDF injection were Negative. Conclusion: The established replication-based real-time quantitative real-time PCR detection method with SIV is reproducible, sensitive and accurate and can be used for the detection of replication-type SIV in recombinant SIV-hPEDF injection.