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Primers and detection method for LAMP detection of arcanobacterium pyogenes

A technique for the detection of Cryptobacterium pyogenes and a detection method, which is applied in the field of primers and detection for the detection of Cryptobacterium pyogenes LAMP, which can solve the problems of inconvenient detection and long time, and achieve the effects of short time-consuming, strong specificity, and low requirements

Inactive Publication Date: 2014-07-23
SHAANXI INST OF ZOOLOGY NORTHWEST INSTOF ENDANGERED ZOOLOGICAL SPECIES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional bacterial culture and biochemical identification take a long time, and PCR detection requires electrophoresis and PCR equipment, which is not convenient for rapid clinical detection in the field

Method used

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  • Primers and detection method for LAMP detection of arcanobacterium pyogenes
  • Primers and detection method for LAMP detection of arcanobacterium pyogenes
  • Primers and detection method for LAMP detection of arcanobacterium pyogenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: The composition of the Cryptobacter pyogenes LAMP detection kit based on the above primers:

[0057] Including the primers mentioned in Table 1 (F3, B3, FIP, BIP, LoopF ​​and LoopB), all synthesized by Invitrogen; 10×Thermopol buffer, dNTPs (10mM), betaine (5M), MgSO4 (100mM), Bst DNA polymerase, chromogenic solution 1000×SYBR green I were purchased from Invitrogen

[0058] Negative control: DEPC water;

[0059] Positive control: the nucleic acid extracted from the standard strain of Cryptobacterium pyogenes ATCC19411 was used as a positive control, and 1 mL of the bacterial enrichment solution was extracted with a promega genomic DNA extraction kit as a template, with a nucleic acid concentration of about 70 ng / uL.

Embodiment 2

[0060] Example 2: Specificity and Sensitivity Test of Detecting Cryptobacter pyogenes Kit

[0061] 1. Detection specificity analysis

[0062] The strains listed in Table 2 were tested by LAMP respectively, and the results showed that the amplified product of the standard strain of Cryptobacter pyogenes was yellow-green after being reacted with SYBR green I (see Figure 2A , of which only the No. 2 detection tube had a color change, while the other detection tubes had no color change), and a typical ladder-like amplification band appeared after agarose gel electrophoresis (see Figure 2B , only No. 2 test tube had a ladder-like amplification band, while the other test tubes had no ladder-like amplification band), and the non-specific amplification of other strains indicated that this method had good specificity for detecting C. pyogenes.

[0063] Table 2 list of experimental strains

[0064] serial number

strain name

Numbering

1

Cryptobacter pyoge...

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Abstract

The invention discloses primers and a detection method for LAMP (Loop-Mediated Isothermal Amplification) detection of arcanobacterium pyogenes, and specifically relates to a visual arcanobacterium pyogenes LAMP detection method which is established by designing LAMP primers according to the PLO gene sequence of the A. pyogenes and is applicable to clinical sample detection, high in specificity and high in sensitivity. The recognition of six specific sequence regions of a target sequence by using six primers designed and screened guarantees high specificity of LAMP; due to the addition of a loop primer, the amplification efficiency is greatly improved and short time is taken.

Description

technical field [0001] The invention belongs to the technical field of cryptobacterium pyogenes detection, and relates to a primer and a detection method for cryptobacterium pyogenes LAMP detection. Background technique [0002] Arcanobacterium pyogenes (A.pyogenes) is a pleomorphic, non-motile Gram-positive bacillus, which was once considered to be Corynebacterium pyogenes, and then classified into the genus Actinomyces. Ramos reclassified it into the genus Cryptobacter after performing phylogenetic analysis based on the 16S rRNA gene. Cryptobacterium pyogenes often parasitizes mucous membranes such as the upper respiratory tract and digestive tract. Such as pneumonia, mastitis, endometritis, arthritis, otitis externa, cystitis and liver, kidney abscess and other symptoms. Tang Jie et al. reported that the infection of musk deer with purulent cryptic rods can cause abscesses in the submandibular and nasal mirrors. And mixed with Escherichia coli, streptococcus, etc., the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6844C12Q2531/119
Inventor 唐婕刘二龙卢丽夏健刘文华胡罕
Owner SHAANXI INST OF ZOOLOGY NORTHWEST INSTOF ENDANGERED ZOOLOGICAL SPECIES
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