炭疽高发地区土壤样本中常见芽胞杆菌的分离及鉴定

Isolation and identification of bacillus species from soil samples in anthrax epidemic area

目的通过分离鉴定炭疽可疑污染土壤样本的芽胞杆菌,评价消毒效果和了解监测地区土壤中的芽胞杆菌分布情况。方法采集炭疽监测点土壤样本60份,对炭疽芽胞杆菌和其他芽胞杆菌进行分离培养和聚合酶链反应扩增鉴定。结果在可疑污染土壤样本中未分离到炭疽芽胞杆菌;从分离到的48个单克隆菌落中扩增到33个目的片段,经测序和blast比对,确定得到地衣芽胞杆菌13株,枯草芽胞杆菌8株,短小芽胞杆菌扩增11株,蜡样芽胞杆菌扩增到1株,巨大芽胞杆菌引物和环状芽胞杆菌引物特异性不好。结论本研究提示该监测点炭疽疫情消毒效果可信,但需要进一步研究验证;几种芽胞杆菌在土壤中广泛存在,在炭疽监测工作中进行病原体分离时需要加以鉴别,可通过特异基因扩增来辅助检验。

Objective To isolate and identify bacillus species in soil samples in anthrax epidemic area and evaluate the disinfection effect and understand the distributions of common bacillus species in the area.Methods Sixty soil samples were collected from the anthrax epidemic area to isolate and identify related bacillus species with PCR assay.ResultsNo Bacillus anthracis was identified,but 13 strains of Bacillus licheniformis,8 strains of Bacillus subtilis,11 strains of Bacillus pumilus and 1 strain of Bacillus cereus were identified in 33 gene fragments from 48 clones by sequencing and blast alignment.The specificities of the primers for Bacillus megatherium and Bacillus circulans were not high.Conclusion The disinfection effect in the anthrax epidemic area was good.The related bacillus species exist widely in the soil,suggesting that their identifications are needed in anthrax surveillance by specific genes amplification