Okumura Takekazu, Nagai Fumiko, Yamamoto Shuta, Oomura Hiroshi, Inouye Kiyoshi, Ito Masahiko, Sawada Haruji
Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, Tokyo 186-8650, Japan.
J Virol Methods. 2005 Mar;124(1-2):143-8. doi: 10.1016/j.jviromet.2004.11.017. Epub 2004 Dec 22.
A simple reverse passive latex agglutination (RPLA) method for detecting white spot syndrome virus (WSSV) in the hemolymph of infected Kuruma shrimp (Penaeus japonicus) was developed. It was confirmed that WSSV could be detected from the shrimp hemolymph when the latex particles blocked with a casein protein were used as detection reagent. It became clear from the result of the infection trial that viruses are detectable by RPLA before the appearance of overt symptoms of this disease. In addition, an amplification product of 982 bp (s) derived from WSSV by PCR was detected in all the samples in which WSSV was detected by RPLA. This newly developed RPLA assay can examine many samples in a simple manner since hemolymph can be extracted more easily than any other organs. This assay can be used conveniently for virus detection in the culture pond of shrimps or in the field.
开发了一种简单的反向被动乳胶凝集(RPLA)方法,用于检测感染的日本对虾(Penaeus japonicus)血淋巴中的白斑综合征病毒(WSSV)。当使用用酪蛋白包被的乳胶颗粒作为检测试剂时,证实可以从虾血淋巴中检测到WSSV。从感染试验结果可以看出,在该疾病出现明显症状之前,通过RPLA可以检测到病毒。此外,在所有通过RPLA检测到WSSV的样品中,均检测到通过PCR扩增得到的来自WSSV的982 bp扩增产物。这种新开发的RPLA检测方法可以以简单的方式检测许多样品,因为血淋巴比任何其他器官更容易提取。该检测方法可方便地用于虾养殖池塘或野外的病毒检测。