Volume 13 Issue 6
Dec.  2017
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CHEN Tiantian, FANG Wenhong, WANG Yuan, LI Xincang, ZHAO Shu, ZHOU Junfang. Establishment and application of a SYBR Green I real-time qPCR detection of Ameson portunus[J]. South China Fisheries Science, 2017, 13(6): 22-29. DOI: 10.3969/j.issn.2095-0780.2017.06.003
Citation: CHEN Tiantian, FANG Wenhong, WANG Yuan, LI Xincang, ZHAO Shu, ZHOU Junfang. Establishment and application of a SYBR Green I real-time qPCR detection of Ameson portunus[J]. South China Fisheries Science, 2017, 13(6): 22-29. DOI: 10.3969/j.issn.2095-0780.2017.06.003
shu

Establishment and application of a SYBR Green I real-time qPCR detection of Ameson portunus

  • 1.

    Key Lab. of East China Sea Fishery Resources Exploitation, Ministry of Agriculture; East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai 200090, China

  • 2.

    College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China

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  • Received Date: February 11, 2017
  • Revised Date: April 12, 2017
  • Accepted Date: May 03, 2017
    Graphical Abstract
    • Abstract

  • Ameson portunus is one of the common pathogens of the swimming crab (Portunus trituberculatus). In this study, a SYBR Green I real time qPCR method whose amplification efficiency reached up to 101.45% with a detection limit as low as 2.0×101 copies·μL-1 was established by targeting A.portunus SSU rRNA gene. The method was A.portunus-specific with good repeatability within and between groups. Then the qPCR method was used to detect farmed swimming crabs from a "toothpaste disease" epidemic area, and the detection rate of A.portunus was 82.35%. The results show that the average muscle load per milligram of the live crabs was lower than that of the dead crabs significantly, which indicates that the death of crabs in the region is related with infection of A.portunus closely. When the load of A.portunus was 1.0×109 copies·mg-1 in the muscle tissue, P.trituberculatus had a risk of illness or even death. Besides, the detection rate for the same samples by a previously established nested-PCR method was only 64.71%, showing that the qPCR method in the present study had higher sensitivity and can provide good technical support for the early diagnosis and prevention of "toothpaste disease".

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