Abstract:
Infectious hypodermal and hematopoietic necrosis virus(IHHNV), as one of the main pathogens of shrimp diseases, can infect many kinds of shrimp, especially juvenile shrimp. Although the mortality rate of the disease is not high, it can cause slow growth of shrimp, resulting in huge economic losses, and seriously affect the sustainable and healthy development of shrimp farming industry. Six LAMP specific primers were designed by Primer Exploer V4 software according to the conservative sequence of IHHNV gene. And then, we had established a rapid detection method for IHHNV which was based on loop-mediated isothermal amplification(LAMP). The sensitivity and specificity of LAMP detection method in this study were analyzed and compared with quantitative real-time PCR(qRT-PCR) and PCR. The results showed that the LAMP method could complete the reaction within 60 minutes under the constant temperature of 63℃, and the positive results were visualized green, while the negative results did not change in color. The minimum detection limit of LAMP method was 10.3 copies/μL. The sensitivity of LAMP method was similar to that of qRT-PCR, which was higher than that of PCR. 50 samples of
Penaeus vannamei collected from different farms were detected by LAMP and PCR respectively. The results of the two methods were consistent. The results showed that the LAMP method was suitable for rapid detection of IHHNV in shrimp.