Primers and method for rapidly identifying group B streptococcus and detecting drug resistance thereof and primer application

A streptococcus and drug resistance technology, which is applied in the field of rapid identification of group B streptococcus and detection of its drug resistance primers and methods and primer application fields, can solve the problems of low sensitivity, complex environment, low specificity, etc., and achieve specificity Strong, sensitive effects

Inactive Publication Date: 2015-01-28
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are many types of microbial flora in the reproductive tract of pregnant women and the environment is complex. The traditional GBS detection method has low specificity and low sensitivity.
In the prior art, the detection method for group B streptococcus mainly relies on the conventional bacteriological culture method, and the detection generally takes 2 to 3 days, which has the disadvantages of cumbersome operation and long time-consuming; the drug resistance detection of group B streptococcus Relying on the results of the drug-sensitive paper test, this method takes a long time to test, and is greatly affected...

Method used

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  • Primers and method for rapidly identifying group B streptococcus and detecting drug resistance thereof and primer application
  • Primers and method for rapidly identifying group B streptococcus and detecting drug resistance thereof and primer application
  • Primers and method for rapidly identifying group B streptococcus and detecting drug resistance thereof and primer application

Examples

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Embodiment 1

[0068] Primers for rapid identification of group B streptococci and detection of their drug resistance, including a primer pair set consisting of four pairs of specific primers;

[0069] Among them, the four pairs of specific primers are: cfb gene primer pair cfb-F and cfb-R for identifying group B streptococcus; ermB gene primer for detecting group B streptococcus resistance to macrocyclic esters For ermB-F and ermB-R; mefA / E gene primer pair mefA / E-F and mefA / E-R for detection of group B streptococcus resistance to macrocyclic esters; for detection of group B streptococcus resistance to linco Amide-resistant linB gene primer pair linB-F and linB-R. Among them, macrocyclic esters, such as erythromycin; lincosamides, such as clindamycin;

[0070] In this embodiment, the gene sequences of four pairs of specific primers are as follows:

[0071] cfb-F: 5'-TTT CAC CAG CTG TAT TAG AAG TA-3'

[0072] cfb-R: 5'-GTT CCC TGA ACA TTA TCT TTG AT-3'

[0073] ermB-F: 5'-GAA AAG GTA CTC...

Embodiment 2

[0080] The method for quickly identifying group B streptococci and detecting its drug resistance is to utilize the primers for quickly identifying group B streptococci and detecting its drug resistance in Example 1 to quickly identify group B streptococcus and detect its drug resistance method, which includes the following steps:

[0081] Step 1, DNA extraction: use the MiniBEST Bacteria Genomic DNA Extraction Kit Ver.3.0 Bacterial Nucleic Acid Extraction Kit to extract the DNA of Group B Streptococcus; and use a nucleic acid protein content analyzer to measure the concentration of the extracted DNA;

[0082] Step 2, primer design: According to the cfb gene sequence of group B streptococci, the ermB gene sequence and mefA / E gene sequence of macrocyclic ester drug resistance, and the linB gene sequence of lincosamide drug resistance collected in GeneBank, Utilize Primer software and Oligo6.0 software to carry out the design of specific PCR primer sequence; The gene sequence of ...

Embodiment 3

[0102] The method for quickly identifying group B streptococci and detecting its drug resistance is to utilize the primers for quickly identifying group B streptococci and detecting its drug resistance in Example 1 to quickly identify group B streptococcus and detect its drug resistance method, which includes the following steps:

[0103] Step 1, DNA extraction: use the MiniBEST Bacteria Genomic DNA Extraction Kit Ver.3.0 Bacterial Nucleic Acid Extraction Kit to extract the DNA of Group B Streptococcus; and use a nucleic acid protein content analyzer to measure the concentration of the extracted DNA;

[0104] Step 2, primer design: According to the cfb gene sequence of group B streptococci, the ermB gene sequence and mefA / E gene sequence of macrocyclic ester drug resistance, and the linB gene sequence of lincosamide drug resistance collected in GeneBank, Utilize Primer software and Oligo6.0 software to carry out the design of specific PCR primer sequence; The gene sequence of ...

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Abstract

The invention relates to the biotechnical field, in particular to primers and a method for rapidly identifying group B streptococcus and detecting drug resistance thereof and primer application. Four pairs of specific primers are as follows: a cfb gene primer pair cfb-F and cfb-R for identifying the group B streptococcus; an ermB gene primer pair ermB-F and ermB-R for detecting the drug resistance of the group B streptococcus to macrocyclic lactones; an mefA/ E gene primer pair mefA/E-F and mefA/E-R for detecting the drug resistance of the group B streptococcus to macrocyclic lactones; an linB gene primer pair linB-F and linB-R for detecting the drug resistance of the group B streptococcus to lincosamides. The method comprises the following steps: Step 1, DNA extraction; Step 2, primer design; Step 3, gene amplification; Step 4, result interpretation. According to the invention, the primers can identify GBS (group B streptococcus) in a genital tract of a pregnant woman and detect the drug resistanceof the GBS to erythromycin and clindamycin at the same time so as to guide clinical medication.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer and a method for rapidly identifying group B streptococcus and detecting its drug resistance and the application of the primer. Background technique [0002] Group B Streptococci (GBS), also known as Streptococcus agalactiae, can cause neonatal pneumonia, meningitis, sepsis and other diseases. Newborns with GBS infection are closely related to GBS colonization of the mother's reproductive tract. Guidelines from the Centers for Disease Control and Prevention (CDC) recommend screening for GBS colonization and antibiotic prophylaxis for pregnant women at 35-37 weeks of gestation. Penicillin is the first choice drug. According to literature reports, 12% of pregnant women are allergic to penicillin. If the pregnant women allergic to penicillin are sensitive to clindamycin drug sensitivity test, the US CDC recommends clindamycin as an alternative drug for pregnant women allergic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12N15/11C12R1/46
CPCC12Q1/689C12Q1/6844C12Q2600/106C12Q2600/16
Inventor 杨维青张丽华何其励张丽王欣吴志毅陈军剑唐小梅郑碧英王佳敏梁有才吴明雨
Owner GUANGDONG MEDICAL UNIV
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