Jupp P G, Grobbelaar A A, Leman P A, Kemp A, Dunton R F, Burkot T R, Ksiazek T G, Swanepoel R
National Institute for Virology, Department of Virology, University of the Witwatersrand, Private Bag X4, Sandringham, Johannesburg 2131, South Africa.
J Med Entomol. 2000 May;37(3):467-71. doi: 10.1093/jmedent/37.3.467.
A reverse transcription-polymerase chain reaction (RT-PCR) was assessed in laboratory tests to detect the presence of single Aedes aegypti (L.) or Eretmapodites quinquevittatus Theobald mosquitoes infected with Rift Valley fever virus in pools of mosquitoes, 50-600 in size, from laboratory colonies or mixed field collections. The viral RNA was detected in all pools containing infected mosquitoes and was shown to be as sensitive as infant mice but more sensitive than Vero cell cultures for virus detection. Pools diluted down to the equivalent of 1:16 000 mosquitoes were also positive by RT-PCR. RNAs from 4 other phleboviruses were negative, there were no false positives and the procedure followed, with the 2 particular primers chosen, gave consistently clear bands of the PCR products on agarose gels without nested PCR being necessary.
在实验室检测中评估了逆转录聚合酶链反应（RT-PCR），以检测来自实验室菌落或混合野外采集的、大小为50至600只的蚊群中感染裂谷热病毒的单个埃及伊蚊（L.）或五带埃蚊（Theobald）蚊子的存在情况。在所有含有感染蚊子的蚊群中均检测到病毒RNA，结果表明其对病毒检测的敏感性与乳鼠相当，但比Vero细胞培养更敏感。稀释至相当于1:16 000只蚊子的蚊群经RT-PCR检测也呈阳性。来自其他4种白蛉病毒的RNA呈阴性，无假阳性，并且按照所采用的程序，选用的2种特定引物在琼脂糖凝胶上始终能产生清晰的PCR产物条带，无需进行巢式PCR。
