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Primer, kit and method for detecting Ichthyophthirius multifiiis

A technology of multi-seed melon worms and kits, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problem of high environmental requirements for storage and use, and rapid detection and diagnosis , did not develop more rapid and simple problems, and achieved the effects of short detection time, high specificity, and enhanced specificity

Inactive Publication Date: 2015-06-17
INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These instruments and equipment are not only of high value, but also have high environmental requirements for their storage and use, and are not suitable for placement and application in aquaculture environments.
At the same time, the common PCR detection method and fluorescent PCR detection method, the detection process is more complicated, can not achieve the purpose of rapid detection and diagnosis
At present, in addition to the development of common PCR and fluorescent PCR methods suitable for the laboratory environment detection and diagnosis of polyzygous melon worms at home and abroad, there has not been developed a more rapid and simple polyzygous insect that is suitable for extensive environments such as aquaculture. Detection and diagnosis technology of small melon worm

Method used

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  • Primer, kit and method for detecting Ichthyophthirius multifiiis

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Embodiment 1

[0033] Example 1: Detection of polyzygos melon beetle

[0034] 1) DNA extraction: Take the scraped body surface of the fish to be tested as a sample, and extract DNA according to the DNA kit method or the classic DNA phenol-chloroform extraction method as the DNA template for LAMP detection. The DNA from the surface scrapings of fish known to be infected with G. multisporidia was used as a positive control, and the DNA from the surface scrapings of known healthy fish was used as a negative control.

[0035] 2) Primer design

[0036] Design of LAMP primers for the polyzygotic melon beetle

[0037] Based on the sequencing results of the 18S rDNA (Sequence ID: gb|DQ270016.1|) of the polyzygotic melon worm as a template, the following 4 primers were designed, and the sequences are as follows:

[0038]3×GCF3:5'-GAT TAT GTC CCT GCC GTT-3' sequence SEQ ID NO:1

[0039] 3×GCB3:5'-TCC TCC TCC TAA ATA AAG TAC A-3' sequence SEQ ID NO:2

[0040] 3×GCFIB: 5’-ACT TCC CAA AGC CTT GCG ...

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Abstract

The invention proposes a primer, a kit and a method for detecting Ichthyophthirius multifiiis. The primer sequence is formed by four sequences in SEQ ID NO:1-4. The invention also protects the kit and the method for detecting the Ichthyophthirius multifiiis. LAMP rapid detection is carried out by using the primer, so that the primer has high sensitivity, strong specificity and short detection time, the detection result can be obtained within about an hour, the detection time is shortened by 2-4 hours in comparison with that of common PCR detection, the operation is simple, and the result is obvious. The overall detection process does not relate to complicated apparatus and equipment, detection can be finished by general technical personnel, the result is easily distinguished, the result can be observed by naked eyes through different colors, fussy electrophoresis and ultraviolet observation are not required, and the primer and the kit are safe.

Description

technical field [0001] The present invention relates to the field of biotechnology. More specifically, it relates to a primer, a kit and a method for detecting the polyzygotic melon worm. Background technique [0002] The many-child melon worms parasitic on freshwater fishes belong to the phylum Protozoa (Portozea), the class Ciliata, the order Holotricha, the family Ophryoglenidae, and the genus Cucurbita ( Ichthyophthirius ). At present, there is only one species of polyzygotic melon beetle ( I. multifiiis ). Small melon bugs are found in tropical, subtropical, and temperate regions, and can extend as far north as the Arctic Circle. [0003] PCR detection technology is a technology for detecting and identifying organisms based on their specific gene fragments. This method has been widely used in the detection of pathogens in humans, animals and plants, and has many advantages such as strong specificity, high sensitivity, and convenience. Sun et al. (2006) reported a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6844C12Q2531/119
Inventor 郭书林
Owner INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU