The quantitative characteristics of infection with infectious precocity virus (IPV) revealed with a new TaqMan probe-based real-time RT-PCR method
文献类型： 外文期刊
作者： Wang, Guohao 1 ; Guo, Xiaomeng 1 ; Huang, Xiao 1 ; Wang, Dehao 1 ; Chen, Yujin 1 ; Qin, Jiahao 1 ; Yang, Guoliang 6 ; Tang, Kathy F. J. 1 ; Dong, Xuan 1 ; Huang, Jie 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Qingdao 266071, Peoples R China
2.Pilot Natl Lab Marine Sci & Technol Qingdao, Lab Marine Fisheries Sci & Food Prod Proc, Qingdao 266071, Peoples R China
3.Minist Agr & Rural Affairs, Key Lab Maricultural Organism Dis Control, Qingdao 266071, Peoples R China
4.Qingdao Key Lab Mariculture Epidemiol & Biosecur, Qingdao 266071, Peoples R China
5.Qingdao Agr Univ, Qingdao 266109, Peoples R China
6.Huzhou Univ, Huzhou 313000, Peoples R China
7.Shandong Agr Univ, Tai An 271000, Peoples R China
8.Dalian Ocean Univ, Dalian 116023, Peoples R China
9.Shanghai Ocean Univ, Shanghai 201306, Peoples R China
10.Jiangsu Shufeng Prawn Breeding Co LTD, Gaoyou 225600, Peoples R China
11.Network Aquaculture Ctr Asia Pacific, Bangkok 10090, Thailand
12.106 Nanjing Rd, Qingdao 266071, Shandong, Peoples R China
关键词： Infectious precocity virus; iron prawn syndrome; TaqMan probe; Real-time RT-qPCR; Macrobrachium rosenbergii
期刊名称：AQUACULTURE （ 2022影响因子：4.5； 五年影响因子：4.6 ）
ISSN： 0044-8486
年卷期： 2023 年 566 卷
收录情况： SCI
摘要： Infectious precocity virus (IPV) is a recently reported RNA virus associated with the sexual precocity of Mac-robrachium rosenbergii, so-called iron prawn syndrome (IPS), and has caused huge economic losses in the prawn farming industry. We developed a semiquantitative approach for diagnosing IPS based on clinical signs and established a one-step, TaqMan probe-based reverse-transcription quantitative PCR (TaqMan-RT-qPCR) method to detect and quantify IPV. The detection limit of this method was as low as 1.00 x 100 copy/reaction. The standard curve between 1.00 x 100 to 1.00 x 109 copies/reaction showed a high correlation coefficient (R2 = 0.999). Quantitative detection of different tissues in M. rosenbergii infected with IPV revealed the characteristic neurotropism of the virus with the highest loads in eyestalks and the brain. The assay showed that the risk of IPS in a farm might be significant if its prawn samples have a load of IPV copies above 103 copies/mu g-RNA. Most symptomatic cases with typical IPS clinical signs had a high load of IPV above 3 x 105 copies/mu g-RNA in eyestalk or 2 x 103 copies/mu g-RNA in cephalothorax tissues. The results showed that TaqMan-RT-qPCR was about 13 and 1300 times more sensitive than reported nested RT-PCR and first-step RT-PCR. Bayesian estimation based on Markov Chain Monte Carlo (MCMC) showed that the TaqMan-RT-qPCR method has the highest diagnostic sensitivity (DSe), and the clinical diagnosis method has the highest diagnostic specificity (DSp). This study developed a specified clinical diagnosis method for IPS and a rapid and sensitive method for IPV detection in farmed prawns and revealed the quantitative characteristics of infection with IPV in M. rosenbergii and provided quantitative statistical evidence supporting IPV being the causative agent of IPS.
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