Streptococcus agalactiae amplification primer group, application and detection method of streptococcus agalactiae
A technology for amplifying primers and Streptococcus lactis, which is applied in the field of microbiology and molecular biology, can solve the problems of designing MCDA amplification primers for Streptococcus agalactiae, and achieve the effect of increasing the amount of MCDA products and the method is convenient
Active Publication Date: 2020-12-18
十堰市太和医院(湖北医药学院附属医院)
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Problems solved by technology
[0006] At present, there is no research on designing MCDA amplification primers for specific genes of Streptococcus agalactiae and combining LFB technology for specific detection
Method used
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Embodiment
[0034] 1. Genome Extraction
[0035] Genomic DNA of Streptococcus agalactiae and other bacteria was extracted using the DNA extraction kit of Tiange Biotechnology Company, and the operation was performed according to the instructions. Determination of the concentration and purity of genomic DNA by UV spectrophotometer, DNA of Streptococcus agalactiae with ddH 2 O serial dilution (from 3ng, 300pg, 30pg, 3pg, 300fg, to 3fg / microliter). A small amount of various genomic DNAs were aliquoted and stored at -20°C for future use.
[0036] 2. Primer Design
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Abstract
The invention relates to a streptococcus agalactiae amplification primer group, an application and a detection method of streptococcus agalactiae, and belongs to the technical field of microbiologicaldetection. The detection method of streptococcus agalactiae comprises the following steps: 1) designing an MCDA amplification primer group of specific genes groEL: CP1, CP2, F1, F2, D1 *, D2, C1, C2,R1 * and R2; 2) in a reaction system comprising the amplification primer group, taking a genome of a streptococcus agalactiae sample to be detected as a template, and performing MCDA constant-temperature amplification to obtain an amplification product; and 3) detecting the amplification product in the step 2) by using a gold nano biosensor to obtain a detection result. According to the method, on the basis of multi-cross constant-temperature amplification and in combination with a nano-biological detection technology, the streptococcus agalactiae can be directly, quickly, sensitively and highly-specifically detected and identified from clinical samples.
Description
【Technical field】 [0001] The invention relates to the technical fields of microbiology and molecular biology, in particular to a Streptococcus agalactiae amplification primer set and its application and a detection method for Streptococcus agalactiae. 【Background technique】 [0002] Streptococcus agalactiae (S. agalactiae) is a β-hemolytic, Gram-positive bacterium. The pathogen can colonize the vagina, intestinal tract and urethra of pregnant women, causing urinary tract infection, bacteremia, chorioamnionitis and placental abruption in pregnant women, which may further induce premature birth and stillbirth, and may also lead to neonatal pneumonia , sepsis and meningitis. Therefore, prenatal screening and early diagnosis of Streptococcus agalactiae in pregnant women can help prevent the occurrence of serious illness in infants. [0003] At present, the detection of Streptococcus agalactiae mainly relies on traditional isolation culture and biochemical identification. This...
Claims
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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/6804C12Q1/04C12N15/11C12R1/46
CPCC12Q1/689C12Q1/6844C12Q1/6804C12Q2531/119C12Q2563/131C12Q2565/607Y02A50/30
Inventor 王梅芳唐以军程雪琴窦志茜杨晶顾玉龙刘得玺谢玲任涛刘岩喻志芳
Owner 十堰市太和医院(湖北医药学院附属医院)