AUTHOR=Wang Yuan , Zhou Jinyang , Yin Menghe , Ying Na , Xiang Yang , Liu Wenchang , Ye Junqiang , Li Xincang , Fang Wenhong , Tan Hongxin 

TITLE=A modification of nested PCR method for detection of Enterocytozoon hepatopenaei (EHP) in giant freshwater prawn Macrobrachium rosenbergii

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=12

YEAR=2022

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2022.1013016

DOI=10.3389/fcimb.2022.1013016

ISSN=2235-2988

ABSTRACT=<p>The microsporidian <italic>Enterocytozoon hepatopenaei</italic> (EHP) has become a critical threat to the global shrimp aquaculture industry, thus necessitating early detection by screening. Development of a rapid and accurate assay is crucial both for the active surveillance and for the assessment of shrimp with EHP infection. In the present study, a distinct strain of <italic>E</italic>. <italic>hepatopenaei</italic> (EHP<italic><sub>Mr</sub></italic>) was found in <italic>Macrobrachium rosenbergii</italic>. The SWP1 gene analysis revealed it was a new genotype that differed with the common strain isolated from the <italic>Litopenaeus vannamei</italic> (EHP<italic><sub>Lv</sub></italic>). A nested SWP-PCR method was modified to fix the bug that the original inner primers could not recognize the EHP<italic><sub>Mr</sub></italic> strain. The redesigned inner primers successfully amplified a product of 182 bp for both the EHP<italic><sub>Mr</sub></italic> strain and the EHP<italic><sub>Lv</sub></italic> strain. The new primers also had good specificity and high sensitivity, which may serve as an alternative for EHP genotyping. This study provided a method for detection of EHP in the biosecurity of <italic>Macrobrachium rosenbergii</italic> farming, and the developed protocol was proposed for the routine investigation and potential carrier screening, especially for molecular epidemiology.</p>