Primers, probes, kit and method for group B streptococci fluorescence quantitative PCR detection
A fluorescence quantitative and streptococcus technology, which is applied in biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of false negative detection, sampling error, sampling failure, etc., and achieve reliable and shortened results. Detection time, effect of excluding false positive results
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Embodiment 1
[0027] Example 1 Fluorescent quantitative PCR detection kit for Group B Streptococcus.
[0028] The group B Streptococcus fluorescent PCR detection kit of this embodiment includes a PCR amplification reaction solution, an enzyme mixture, a positive control and a negative control.
[0029] Wherein said PCR amplification reaction liquid mainly comprises:
[0030] ① PCR buffer solution (PCR Buffer), composed of tris-HCl, KCl and purified water to provide the pH value and salt ion environment required for the reaction;
[0031] ② MgCl 2 , providing a catalyst for the reaction enzyme;
[0032] ③ dNTPs, which provide the raw materials required for the formation of DNA chains during the reaction;
[0033] ④ Detection of group B streptococci and human β-actin primer pairs and corresponding probes.
[0034] The primer pair GBS-F, GBS-R and probe GBS-P as described in table 1 are primers and probes derived from the conserved region of group B streptococci; primer pair β-actin-F, β-a...
Embodiment 2
[0037] Example 2 Detection of Group B Streptococcus.
[0038] Step A, extract sample DNA and prepare DNA template.
[0039] The anal swab and vaginal swab samples from 12 cases from the hospital that were negative in culture test and 10 cases were positive in culture test were used to extract nucleic acid and prepare DNA template according to the kit instructions. In this example, Jifan Biotechnology ( The FinePure virus DNA / RNA column extraction kit provided by Beijing) Co., Ltd. is operated according to the instructions of the kit, and other DNA extraction methods in the prior art can also be used.
[0040] Step B, adding samples: add the 22 sample DNAs prepared in step A and the positive control and negative control in the kit respectively to the PCR tubes containing the PCR amplification reaction solution to obtain the corresponding sample PCR reaction system, positive PCR reaction system and negative PCR reaction system.
[0041] The composition of the PCR reaction syst...
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