Primer for detecting reverse transcription fluorescence PCR (polymerase chain reaction) of salmonella paratyphi A

A technology for paratyphoid and Salmonella, which is applied in the field of primers for reverse transcription fluorescent PCR detection, can solve the problems of unfavorable early diagnosis, low positive rate, time-consuming H-a antigen, etc.

Active Publication Date: 2012-08-15
ICDC CHINA CDC
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Problems solved by technology

These methods still have defects such as relatively low positive rate, time-consuming induction of H-a antigen, and unfavorable early diagnosis, which brings certain difficulties to epidemiological tracking, early control and early clinical diagnosis. Therefore, it is necessary to establish A Rapid and Accurate Molecular Biological Detection Method for Salmonella

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  • Primer for detecting reverse transcription fluorescence PCR (polymerase chain reaction) of salmonella paratyphi A
  • Primer for detecting reverse transcription fluorescence PCR (polymerase chain reaction) of salmonella paratyphi A
  • Primer for detecting reverse transcription fluorescence PCR (polymerase chain reaction) of salmonella paratyphi A

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Embodiment

[0014] The design and detection method of the reverse transcription fluorescent PCR detection primer of embodiment Salmonella paratyphi A

[0015] 1 Materials and methods

[0016] 1.1 Experimental strains

[0017] 44 strains of Salmonella paratyphi used in the present embodiment, Salmonella typhi, Salmonella typhi B and C and other 31 non-typhoidal Salmonella serotypes total 57 strains (table 1), all serotypes Serotyping was performed using Salmonella antiserum (purchased from Statens Serum Institut, SSI, Denmark). In addition, other common intestinal pathogens and Streptococcus pneumoniae, Borrelia burgdorferi, Leptospira, Legionella pneumophila, Neisseria meningitidis, Rickettsia, Brucella, etc. Bacteria and other bacterial strains (Table 1) were used for specificity and sensitivity evaluation, and the above-mentioned bacterial strains were all from the Institute of Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention.

[0018] 1.2 ...

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Abstract

The invention provides a primer for detecting reverse transcription fluorescence PCR (polymerase chain reaction) of salmonella paratyphi A, which comprises 5'-CCTGTTAGAAGTGTTTACAACTT-3' and 5'-GCTAAACCACCAAATTGTGT-3'. The invention further provides a kit which contains the primer and is used for detecting the salmonella paratyphi A. The primer is a specificity primer which is designed for hsdM genes of the salmonella paratyphi A. In the reverse transcription fluorescence PCR for detecting the target genes, 44 types of salmonella paratyphi A detected by the method are positive, amplification results of other 34 types of non-salmonella typhi blood serum, other 5 types of pathogenic entero bacteria causing diarrhea and 8 types of common non-salmonella mainly causing fever are negative, accordingly, the method is high in specificity and sensitivity, and lower limit of detection of the reverse transcription fluorescence PCR is 5pg / reaction.

Description

technical field [0001] The invention relates to the detection of Salmonella paratyphi A, in particular to primers for reverse transcription fluorescent PCR detection of Salmonella paratyphi A. Background technique [0002] Salmonella paratyphi A (S.Paratyphi A) is one of the most common pathogens in intestinal infectious diseases. The rapid and accurate detection of Salmonella paratyphi A is an important means to control the epidemic of paratyphi A. At present, the main methods for the routine diagnosis of paratyphoid fever in the laboratory are bacterial culture, Widal agglutination reaction, and typhoid blood cell rapid diagnosis method. These methods still have defects such as relatively low positive rate, time-consuming induction of H-a antigen, and unfavorable early diagnosis, which brings certain difficulties to epidemiological tracking, early control and early clinical diagnosis. Therefore, it is necessary to establish Rapid and Accurate Molecular Biological Detectio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12R1/42
Inventor 闫梅英樊粉霞阚飙
Owner ICDC CHINA CDC
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