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Group B streptococcus nucleic acid assay kit based on PCR (polymerase chain reaction) fluorescent probe method

A technology for detecting kits and fluorescent probes, which is applied in the field of group B Streptococcus PCR fluorescent probe method nucleic acid detection kits, can solve the problems of poor detection effect and low sensitivity of kits, and achieve high sensitivity and wide detection range , the effect of quick operation

Inactive Publication Date: 2016-03-02
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The invention provides a group B streptococcus PCR fluorescent probe method nucleic acid detection kit to solve the problems of poor detection effect and low sensitivity of the kit in the prior art

Method used

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  • Group B streptococcus nucleic acid assay kit based on PCR (polymerase chain reaction) fluorescent probe method
  • Group B streptococcus nucleic acid assay kit based on PCR (polymerase chain reaction) fluorescent probe method
  • Group B streptococcus nucleic acid assay kit based on PCR (polymerase chain reaction) fluorescent probe method

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Embodiment 1

[0033] This embodiment provides a nucleic acid detection kit for Group B Streptococcus PCR fluorescent probe method. The nucleic acid detection kit for Group B Streptococcus PCR fluorescent probe method includes a nucleic acid release agent and a PCR reaction solution, and the nucleic acid release agent includes Savantine 0.05-0.5mM / L, potassium chloride 100-300mM / L, sodium dodecylsulfonate 0.05-3% and ethanol 0.01-1%; the PCR reaction solution further includes the following components:

[0034] (1) 0.2 μmol / L-0.5 μmol / L probe for detection of target polynucleotide, the sequence of the probe for detection of target polynucleotide is 5'-CAAATAACATTTCATATGTTATCTGGCAACAAAAGT-3'.

[0035] (2) 0.2mmol / L deoxyribonucleoside triphosphate.

[0036] (3) 0.2μmol / L-0.5μmol / L upstream primers and downstream primers for target polynucleotide amplification, said upstream primers for target polynucleotide amplification, downstream primers and target polynucleotides The probes for acid detec...

Embodiment 2

[0046] This embodiment provides the Group B Streptococcus nucleic acid detection kit described in Example 1 above for detecting Group B Streptococcus-DNA in unknown samples such as genital secretions and rectal secretions. The operation steps are:

[0047] 1. Reagent preparation

[0048] According to the number of samples to be tested, negative control and positive control, take the corresponding amount of reaction solution and enzyme mixture in proportion (reaction solution 38-42 μl / person + enzyme mixture 1-2 μl / person + internal standard 1 μl / person) Solution and internal standard, mixed thoroughly to form a PCR-mix, and centrifuged briefly for later use.

[0049] 2. Sample processing

[0050] 1. Method A: one-step method

[0051] (1) Add 3-5 μl of nucleic acid release agent to each PCR reaction tube (deep suction and shallow pipetting are recommended to avoid air bubbles), and then add 2-4 μl each of the sample to be tested, the negative control, and the positive control...

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Abstract

The invention provides a group B streptococcus nucleic acid assay kit based on a PCR (polymerase chain reaction) fluorescent probe method. The group B streptococcus nucleic acid assay kit comprises nucleic acid releasing agents and PCR reaction fluid, wherein the nucleic acid releasing agents include 0.05-0.5mM / L surfactin, 100-300mM / L potassium chloride, 0.05-3% sodium dodecyl sulfate and 0.01-1% ethyl alcohol, the PCR reaction fluid comprises a forward primer, a reverse primer and a probe, the forward primer and the reverse primer are used for target polynucleotide amplification, and the probe is used for target polynucleotide assay. The group B streptococcus nucleic acid assay kit has the advantages that a nucleic acid release method is adopted, a pathogen coat protein structure is rapidly destroyed by strong protein denaturants, group B streptococcus-DNA (deoxyribonucleic acid) in unknown samples such as genital secretions and rectum secretions of perinatal pregnant women can be assayed rapidly, and assay sensitivity can be up to 500 copies / ml, so that reliable experimental bases are provided for group B streptococcus infection diagnosis, and the problems of poor assay specificity and low sensitivity of the prior art can be solved.

Description

technical field [0001] The invention relates to a group B streptococcus nucleic acid detection kit, in particular to a B group streptococcus PCR fluorescent probe method nucleic acid detection kit. Background technique [0002] Group B streptococcus (group B streptococcus, GBS) is an opportunistic pathogen that parasitizes in the human genitourinary tract and lower gastrointestinal tract, and the carrier rate of healthy people can reach 35%. As early as the 1970s, group B streptococci had been confirmed as one of the important pathogenic bacteria for perinatal maternal and infant infection. Group B streptococcus colonized in the maternal genital tract can be vertically transmitted to newborns during childbirth, and neonatal sepsis, meningitis and pneumonia caused by group B streptococcal infection in newborns have a high fatality rate and can lead to Neurological sequelae. In 2010, the US Centers for Disease Control and Prevention formulated the "Guidelines for Perinatal G...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/46
CPCC12Q1/686C12Q1/689C12Q2563/107C12Q2545/101
Inventor 戴立忠黄河邓中平易晓明
Owner SANSURE BIOTECH INC
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