Comparison of nucleic acid-based detection methods for Ecytonucleospora hepatopenaei in Penaeus vannamei

被引：0
作者：
Yang, Jinyu [1 ,2 ,3 ]
Zhang, Lu [1 ,2 ,3 ]
Han, Lulu [1 ,2 ,3 ]
Liu, Junjiang [1 ,2 ,3 ]
Jia, Xin [1 ,2 ,3 ]
Wang, Mengqiang [1 ,2 ,3 ,4 ]
机构：
[1] Ocean Univ China, MOE, Shandong Key Lab Marine Seed Ind preparatory, Key Lab Marine Genet & Breeding, Qingdao 266003, Peoples R China
[2] Ocean Univ China, Qingdao Inst Maritime Silk Rd, Qingdao Inst Blue Seed Ind, Qingdao 266003, Peoples R China
[3] Ocean Univ China, Sanya Oceanog Inst, Hainan Key Lab Trop Aquat Germplasm, Sanya 572024, Peoples R China
[4] Southern Marine Sci & Engn Guangdong Lab Guangzhou, Guangzhou 511458, Peoples R China
来源：
AQUACULTURE | 2025年 / 600卷
关键词：
Pathogen detection; Enterocytozoon hepatopenaei; Penaeus vannamei; ENTEROCYTOZOON-HEPATOPENAEI; SHRIMP; AMPLIFICATION; EHP;
D O I：
10.1016/j.aquaculture.2025.742218
中图分类号：
S9 [水产、渔业];
学科分类号：
0908 ;
摘要：
Ecytonucleospora hepatopenaei (EHP) has caused significant losses in the global shrimp farming industry. As no effective treatment is available, early diagnosis has become the best option to curb the widespread transmission of EHP. Molecular biological diagnosis has become the gold standard among many diagnostic methods, due to its high sensitivity and specificity. There is an urgent need for a high-performance and low-cost diagnostic method to detect shrimp disease during the incubation period to reduce losses. In this study, we conducted a systematic comparison of 18 recently published and classic molecular biological diagnostic methods, such as polymerase chain reaction (PCR), nested PCR, quantitative real-time PCR (qPCR), loop-mediated isothermal amplification (LAMP), and recombinant polymerase amplification (RPA). This study examined the sensitivity, stability, specificity, and actual detection rate of the different methods. Compared with all the other diagnostic methods, the LAMP-4 method can detect 101 copies/L within 30 min, with a 100 % practical detection rate, which represents a relatively high level of stability and specificity. Moreover, LAMP methods do not require expensive thermal cycling equipment or post-processing of the product, which could help to prevent interference with subsequent testing. When combined with other rapid nucleic acid extraction methods, LAMP methods seem to be more suitable for EHP field detection.
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页数：7
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共 37 条
  • [1] Amornrat Tangprasittipap, Jiraporn Srisala, Saisunee Chouwdee, Et al., The microsporidian Enterocytozoon hepatopenaei is not the cause of white feces syndrome in whiteleg shrimp Penaeus (Litopenaeus) vannamei, BMC veterinary research, 9, pp. 1-10, (2013)
  • [2] Anantasomboon G., Sriurairatana S., Flegel T.W., Withyachumnarnkul B., Unique lesions and viral-like particles found in growth retarded black tiger shrimp Penaeus monodon from East Africa, Aquaculture, 253, 1-4, pp. 197-203, (2006)
  • [3] Aranguren L.F., Han J.E., Tang K.F.J., Enterocytozoon hepatopenaei (EHP) is a risk factor for acute hepatopancreatic necrosis disease (AHPND) and septic hepatopancreatic necrosis (SHPN) in the Pacific white shrimp Penaeus vannamei, Aquaculture, 471, pp. 37-42, (2017)
  • [4] Arunrut N., Tondee B., Khumwan P., Kampeera J., Kiatpathomchai W., Rapid and sensitive colorimetric detection of microsporidian Enterocytozoon hepatopenaei (EHP) based on spore wall protein (SWP) gene using loop-mediated isothermal amplification combined with DNA functionalized gold nanoparticles as probes, Aquaculture, 533, (2021)
  • [5] Boyd C.E., Davis R.P., McNevin A.A., Perspectives on the mangrove conundrum, land use, and benefits of yield intensification in farmed shrimp production: a review, J. World Aquacult. Soc., 53, 1, pp. 8-46, (2022)
  • [6] Cai S.-X., Kong F.-D., Xu S.-F., Yao C.-L., Real-time loop-mediated isothermal amplification for rapid detection of Enterocytozoon hepatopenaei, PeerJ, 6, (2018)
  • [7] Chaijarasphong T., Munkongwongsiri N., Stentiford G.D., Aldama-Cano D.J., Thansa K., Flegel T.W., Sritunyalucksana K., Itsathitphaisarn O., The shrimp microsporidian Enterocytozoon hepatopenaei (EHP): biology, pathology, diagnostics and control, J. Invertebr. Pathol., 186, (2021)
  • [8] Chen W.F., Fu Y.W., Zeng Z.Y., Guo S.Q., Yan Y.L., Tu Y.F., Gou T.G., Zhang Q.Z., Establishment and application of a TaqMan probe-based qPCR for the detection of Enterocytozoon hepatopenaei in shrimp Litopenaeus vannamei, Parasitol. Res., 121, 8, pp. 2263-2274, (2022)
  • [9] Hou Z.-H., Yu J.-Y., Wang J.-J., Li T., Chang L.-R., Fang Y., Yan D.-C., Development of a PCR assay for the effective detection of Enterocytozoon hepatopenaei (EHP) and investigation of EHP prevalence in Shandong Province, China, J. Invertebr. Pathol., 184, (2021)
  • [10] Jaroenram W., Owens L., Separation of endogenous viral elements from infectious Penaeus stylirostris densovirus using recombinase polymerase amplification, Mol. Cell. Probes, 28, 5-6, pp. 284-287, (2014)
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