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| Research of Quantification for IHHNV Pseudovirus Nucleic Acid Reference Materials |
| CHEN Gui-fang1,LI Hui-jie2,GAO Yun-hua3,YANG Jia-yi3,DONG Lian-hua3 |
1. College of Chemistry Engineering, Shenyang University of Chemistry Technology, Shenyang, Liaoning 110142,China
2. Shenzhen Institute for Technology Innovation, National Institute of Metrology, Shenzhen, Guangdong 518107,China
3. Center for Advanced Measurement Science, National Institute of Metrology, Beijing 100029,China |
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Abstract Infectious hypodermal and haematopoietic necrosis virus (IHHNV) infection caused acute infectious diseases, especially to juvenile shrimp. According to the IHHNV gene sequence (AF218266.2) accepted by GenBank, pseudoviruses containing the whole genome sequence of IHHNV have been produced. Referring to GB/T 25878-2010 guidance on IHHNV PCR test, ORF1 gene was aimed to design special primers and probes, digital PCR detection method for IHHNV was established. Then digital PCR method was employed to verify the uniformity and stability of the reference material. The reference values with expanded uncertainty was (1.22±0.15)×103copies/μL (k=2). Multiple dPCR platforms were recruited to validate the values of the reference materials, and the relative standard deviations value was less than 4%, showing good reproducibility. Commercial IHHNV nucleic acid detection kit was used to detect the reference materials, the results were in good agreement with the reference values.
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Received: 20 July 2022
Published: 08 March 2023
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2023, Vol. 44 
