Establishment And Application Of Fluorescence PCR Detection Technology Of Group B Streptococcal Infection In Pregnant Women

[Background]Group B Streptococcus(GBS) weregram positive cocci, is the primary pathogen perinatal and neonatal infection disease. GBS according to the specificity of the type can be divided into at least 10 serotypes, any serotype can cause early onset of GBS infection. Studies have found that, GBS also can cause urinary tract infections, chorioamnionitis, postpartum endometritis, wound infection, production and / or postpartum bacteremia and stillbirth, reproductive tract GBS colony can cause or preterm premature rupture of membranes, GBS has become the 2 months of bacteremia and meningitis after the birth of the most common pathogenic bacteria.GBS current detection methods are: pathogen isolation, Gram staining, immunofluorescent antibody test, serum starch medium colorimetric method, many kinds of method for detection of antigen as co agglutination test and latex agglutination test and enzyme-linked immunosorbent assay, and DNA probe, but serological methods exist time-consuming and labor-intensive, low accurate rate and while the conventional method for rapid detection of molecular biology, new development in some aspects can make up for the lack of serological, but environmental pollution, reproducibility, sensitivity, high false positive and other shortcomings limit the application and popularization of. [Objective]To establish a reasonable detection method, rapid detection of group B streptococcus infection in pregnant women. [Methods]This study adopts the real-time PCR amplification of specific DNA target fragment, detection and real-time monitoring of PCR products were hybridized using fluorescent labeled probe. Through verification, evaluation of GBS primer probe or get out of danger, the minimum detection linear range, accuracy, specificity, repeatability, interfering substances, using fluorescent PCR method and sequence analysis for the simultaneous detection of samples, the detection results of various detection methods, fluorescent PCR reagent sensitivity, specificity, the total coincidence rate the Kappa value, the hypothesis test and evaluation of consistency of Kappa value(U), the authenticity and reliability evaluation of clinical evaluation test. [Results]The results showed: the sensitivity of the reaction system was determined as 1×104CFU/ml, the linear range of 1×108 copies/ml to 2×103copies/ml, the accuracy rate was 100%, the specificity is good, there is no interference, precision reference variation coefficient of CV was less than 5%, the repeatability meets the requirements. With the establishment of a method for the detection of First Hospital Affiliated to Suzhou University in 200 samples, 37 were positive detected in all, 163 negative. [Conclusions]The performance evaluation test and clinical sample verification, real-time fluorescent quantitative PCR method established in this study is a kind of strong specificity, high sensitivity, good repeatability, fast and secure detection method, can be used in the rapid detection of GBS.