利用聚合酶链反应从环境中检测炭疽芽孢杆菌的孢子。

Detection of spores of Bacillus anthracis from environment using polymerase chain reaction.

作者信息

Alam Syed Imteyaz, Agarwal Gauri Shankar, Kamboj Dev Vrat, Rai Ganga Prasad, Singh Lokendra

机构信息

Division of Biotechnology, Defence R & Establishment, Jhansi Road, Gwalior 474 002, India.

出版信息

Indian J Exp Biol. 2003 Feb;41(2):177-80.

PMID:15255613

Abstract

A sensitive PCR based detection of Bacillus anthracis spores from environnment was standardized. Specific 1247bp amplicon could be detected with template concentration as low as 13 pg. Sensitivity was enhanced to 10 fold by nesting with second set of primers, forming 208bp amplicon. Extraction of DNA from spores purified from soil samples by aqueous polymer two-phase system followed by partial germination and freeze-thaw treatment yielded best results. Soil sample spiked with spores (8x10(2)/g of sample) could be detected with this method.

摘要

一种基于PCR的从环境中灵敏检测炭疽芽孢杆菌孢子的方法被标准化。使用低至13 pg的模板浓度即可检测到特异性的1247 bp扩增子。通过用第二组引物进行巢式PCR，灵敏度提高了10倍，形成了208 bp的扩增子。通过水相聚合物双相系统从土壤样品中纯化的孢子中提取DNA，随后进行部分萌发和冻融处理，可获得最佳结果。用这种方法可以检测到添加了孢子（8×10²/g样品）的土壤样品。