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首页> 外文期刊>Laboratory Animal Science >SCREENING FOR SIMIAN TYPE-D RETROVIRUS INFECTION IN MACAQUES, USING NESTED POLYMERASE CHAIN REACTION
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SCREENING FOR SIMIAN TYPE-D RETROVIRUS INFECTION IN MACAQUES, USING NESTED POLYMERASE CHAIN REACTION

机译：嵌套式聚合酶链反应筛查猕猴中猿猴D型逆转录病毒感染

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摘要

A nested polymerase chain reaction (PCR) assay was developed to detect proviral DNA in peripheral blood mononuclear cells (PBMC) of macaques infected with simian type-D retrovirus (SRV/D). Primers were designed to amplify gag gene sequences of SRV/D serotype 1, 2, and 3 viral genomes and were used in a single assay for simultaneous detection of infection with SRV/D-1, SRV/D-2, or SRV/D-3. Results of plasmid dilution studies indicate sensitivity of nested PCR in the range of I to 10 genomic copies. The PBMC samples from 395 macaques of unknown SRV/D status, obtained from several primate facilities, were tested in parallel by Western blot (immunoblot) analysis, virus isolation, and nested PCR. Infection was detected in 60 (15.2%) animals by nested PCR, in 40 (10.1%) animals by virus isolation, and in 28 (7.1%) animals by immunoblot. All 40 culture-positive samples were positive by nested PCR. in addition, 11 of 23 immunoblot-positive/virus isolation-negative samples, 2 of 20 immunoblot-indeterminate/virus isolation-negative samples, and 7 of 312 immunoblot-negative/virus isolation-negative samples were identified as positive by nested PCR Nested PCR is a sensitive and specific assay for simultaneous screening for infection with serotypes 1, 2, and 3 of simian type D retrovirus, and is a powerful tool for rapid screening and surveillance in macaque colonies.
机译：建立了巢式聚合酶链反应（PCR）检测试剂盒，以检测感染猿猴D型逆转录病毒（SRV / D）的猕猴的外周血单个核细胞（PBMC）中的原病毒DNA。设计引物以扩增SRV / D血清型1、2和3病毒基因组的gag基因序列，并用于单一测定中，以同时检测SRV / D-1，SRV / D-2或SRV / D的感染-3。质粒稀释研究的结果表明巢式PCR的敏感性在1至10个基因组拷贝的范围内。通过Western印迹（免疫印迹）分析，病毒分离和巢式PCR并行测试了来自数个灵长类设施的395只未知SRV / D状态猕猴的PBMC样品。通过巢式PCR在60只（15.2％）动物中检测到感染，通过病毒分离在40只（10.1％）动物中检测到感染，通过免疫印迹在28只（7.1％）动物中检测到感染。巢式PCR显示所有40个培养阳性样品均为阳性。此外，通过巢式PCR巢式PCR检测，在23个免疫印迹阳性/病毒分离阴性样品中，有11个，在20个免疫印迹不确定/病毒分离阴性样品中的2个和在312个免疫印迹阴性/病毒分离阴性样品中的7个被鉴定为阳性PCR是用于同时筛选猿猴D型逆转录病毒1型，2型和3型感染的灵敏且特异的测定方法，是在猕猴菌落中进行快速筛选和监测的有力工具。

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