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Set type PCR method detection of Pinna(Atrina)pectinate pathogeny Rickett's organism

A technology of rickettsia-like rickettsia and chlamydia, which is applied in the determination/testing of microbes, biochemical equipment and methods, etc., can solve the problems that no effective method for rickettsia-like rickettsia has been provided, and achieve sensitive The effect of specificity, strong specificity and high sensitivity

Inactive Publication Date: 2006-11-15
ZHEJIANG UNIV +1
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  • Claims
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Problems solved by technology

[0005] The prior art does not provide an effective method for the detection of Rickettsia-like species that cause morbidity and mortality in Chlamys farreri

Method used

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  • Set type PCR method detection of Pinna(Atrina)pectinate pathogeny Rickett's organism
  • Set type PCR method detection of Pinna(Atrina)pectinate pathogeny Rickett's organism

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Experimental program
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Embodiment Construction

[0037] The result of the embodiment carried out by the technical scheme detection operation step:

[0038] Nested PCR detected 35 scallop samples, and the positive results of nested PCR only appeared in the samples taken from the farms where a large number of deaths occurred in Qingdao and Long Island. Strong; Microscopic examination of these samples mostly showed Rickettsia-like infection, but there were also individual scallops with normal appearance and no obvious symptoms, and no Rickettsia-like infection was found in microscopic examination, but positive bands appeared in PCR detection. The band is weak, but clearly visible (see figure 2 ).

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Abstract

The nested PCR method for detection of the pathogenic Rickettsia-like species of Chlamys farreri mainly solves the early, rapid, sensitive and specific detection of Rickettsia-like species that cause the disease and death of Chlamys farreri. The detection method firstly purifies Rickettsia-like organisms from scallop tissue; then, according to our completed 16S rDNA sequences with conserved functions in bacterial taxonomy, three unique segments of DNA sequence design and synthesis of inner primers and outer primers, the primer sequences are: outer primer: Out1.5'-CTTCTTCGGAGGTGTATTCT-3', Out2.5'-TTGCGACCGTACTCCC-3'; inner primer: In1.5'-AATCCGAAGACCCGACGTTTA-3', In2.5'-GCCAAACTTGAGATCGGAAGA-3'. Prepare the DNA template of Rickettsia spp.; finally, perform nested PCR amplification to detect the pathogen Rickettsia spp. By using the invention for regular sampling detection, Rickettsia-like infection can be found early, which is convenient for early diagnosis of the disease, and measures can be taken in time to prevent the outbreak of the disease.

Description

technical field [0001] The invention belongs to the field of biotechnology of fishery science, namely, the application of modern molecular biology technology—PCR technology to solve the problem of pathogenic detection of rickettsia-like scallops (Chlamys farreri in Latin name) that occurs in seawater culture. The invention is to create a nested PCR (English name is nested PCR) to detect the Rickettsia-like organism (English name is rickettsia-likeprokaryote, RLP or rickettsia-like organism, RLO) that causes the disease and death of Chlamys farreri. )Detection method. Background technique [0002] Scallop farming is an important pillar industry that dominates shellfish farming or even the entire mariculture in my country. Chlamys farreri is an important marine cultured scallop species in China. However, since 1995, scallops have been dying on a large scale, especially in 1997-1999. During the year, the mortality rate of scallops reached more than 90%, and in some sea areas 10...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 吴信忠李登峰
Owner ZHEJIANG UNIV