Quantitative detection method of infectious hypodermal and hematopoietic necrosis virus of penaeus orientalis

Abstract

The invention relates to a quantitative detection method of infectious hypodermal and hematopoietic necrosis virus of penaeus orientalis. The method comprises the following steps: firstly, extractinga gene group DNA of a to-be-detected sample; designing a primer and a TaqMan probe according to related sequences of IHHNV; finally, carrying out fluorescent quantitative PCR (Polymerase Chain Reaction) detection on the gene group DNA of the to-be-detected sample by adopting the designed primer and the TaqMan probe. The detection method disclosed by the invention is high in detection sensitivity and high in specificity, the lowest copy number which can be detected is 7, the time consumption is short, and a situation of false positive is avoided, moreover, the method can be applied to moleculardisease detection and early warning in penaeus orientalis culture in China, and has good application prospect.

Description

technical field [0001] The invention discloses a quantitative detection method for shrimp infectious subcutaneous and hematopoietic tissue necrosis virus, which belongs to the technical field of molecular biology. Background technique [0002] Infectious hypodermic and hematopoietic necrosis virus (IHHNV) is a small single-stranded DNA virus. It is the smallest particle among the known shrimp viruses. Virion has a diameter of about 22nm (nano), nonenveloped, and Icosahedron symmetrical spherical particles. The parts of IHHNV infection in shrimp include Ectodermal tissues such as gills, epidermis, anterior and posterior enterocytes, nerve cords, and ganglia; and mesodermal organs such as hematopoietic tissue, antennal glands, gonads, lymphoid organs, connective tissue, and striated muscles. Inclusion bodies form in the host cell nucleus. In the white prawn (Litopenaeus vannamei), it causes Runt-deformity syndrome (RDS), which grows slowly and unevenly, the frontal angle is ...

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Problems solved by technology

The PCR method has the advantage of high sensitivity, but generally PCR products need to be observed by agarose gel electrophoresis and ethidium bromide staining with ultraviolet light, which not only requires a variety of instruments, but also is time-consuming and labor-intensive, but it is harmful to health

Nested PCR requires two pairs of primers, which takes a long time; the fluorescent quantitative dye method can be known to have the disadvantages of false positives and high cost based on the principle

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