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目的从阴道炎患者的棉拭子样本中分离筛选具有抗菌活性乳酸菌,为研究对细菌性阴道炎具有确实疗效的微生态制剂提供理论与菌种保证。方法应用纯培养技术及双层平板法从阴道样品中分离筛选具有较好抑菌活性的乳酸菌。通过有机酸排除试验、过氧化氢排除试验和蛋白类抑菌物质排除试验对具有较好抑菌活性菌株的主要抑菌物质分析,并通过PCR反应对是否具有细菌素相关基因进行鉴定。结果从7个棉拭子样本中共分离得到26株乳酸菌,其中有6株呈现抑菌活性。K4M2、K4M5、K4M6和K4M8菌株的主要抗菌物质为有机酸。而K4M9、K4M10两株植物乳杆菌的主要抗菌物质除有机酸外,尚可以产生细菌素。病原菌拮抗试验结果表明,这两株菌株的混合培养上清对具有较好的抑菌效果。结论 K4M9、K4M10菌株有望作为微生态制剂用于细菌性阴道炎的临床防治。
OBJECTIVE To isolate and screen lactic acid bacteria with antibacterial activity from cotton swab samples of vaginitis patients and to provide theory and species guarantee for the study of probiotics that have definite curative effect on bacterial vaginosis. Methods Pure culture technique and double-layer plate method were used to separate and screen lactic acid bacteria with good antibacterial activity from vaginal samples. Through the organic acid elimination test, hydrogen peroxide elimination test and protein antibacterial substance elimination test, the main bacteriostatic substances with good antibacterial activity were analyzed, and whether there was bacteriochlorin-related gene was identified by PCR reaction. Results Totally 26 strains of lactic acid bacteria were isolated from 7 swab swabs, of which 6 strains showed antibacterial activity. The main antibacterial substances of K4M2, K4M5, K4M6 and K4M8 strains are organic acids. The K4M9, K4M10 two strains of Lactobacillus plant antibacterial substances in addition to organic acids, but also can produce bacteriocins. The results of pathogen antagonistic test showed that the mixed culture supernatant of the two strains had good bacteriostasis effect. Conclusion K4M9 and K4M10 strains are expected to be used as probiotics for the prevention and treatment of bacterial vaginosis.