Ibrahim M S, Turell M J, Knauert F K, Lofts R S
Diagnostic Systems Division, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702, USA.
Mol Cell Probes. 1997 Feb;11(1):49-53. doi: 10.1006/mcpr.1996.0075.
A reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect Rift Valley fever (RVF) virus RNA in experimentally infected mosquitoes was developed. The specificity of the assay was evaluated with three other phleboviruses; sandfly fever Sicilian (Sabin), sandfly fever Naples (Sabin) and Punta Toro (MSP 3) viruses. The relative sensitivity of the assay, determined by using RVF virus RNA extracted from serial dilutions of virus culture, was approximately 50 plaque forming units. This sensitivity level was 100-fold higher when a nested PCR procedure was used. When the RT-PCR assay was used with coded samples of intrathoracically-infected and uninfected mosquito, the assay detected the virus in all infected mosquitoes. With this assay, it was possible to detect RVF virus RNA in a single infected mosquito in the background of 10, 25 or 50 uninfected mosquitoes.
开发了一种逆转录聚合酶链反应（RT-PCR）检测方法，用于检测实验感染蚊子体内的裂谷热（RVF）病毒RNA。该检测方法的特异性通过另外三种白蛉病毒进行评估，即西西里白蛉热（萨宾株）、那不勒斯白蛉热（萨宾株）和蓬塔托罗病毒（MSP 3）。通过使用从病毒培养物系列稀释液中提取的RVF病毒RNA来确定该检测方法的相对灵敏度，约为50个空斑形成单位。当使用巢式PCR程序时，该灵敏度水平提高了100倍。当将RT-PCR检测方法用于胸腔内感染和未感染蚊子的编码样本时，该检测方法在所有感染蚊子中均检测到了病毒。使用该检测方法，可以在10只、25只或50只未感染蚊子的背景中检测到单个感染蚊子体内的RVF病毒RNA。
