Jackson P J, Hugh-Jones M E, Adair D M, Green G, Hill K K, Kuske C R, Grinberg L M, Abramova F A, Keim P
Environmental Molecular Biology Group, Los Alamos National Laboratory, Los Alamos, NM, USA.
Proc Natl Acad Sci U S A. 1998 Feb 3;95(3):1224-9. doi: 10.1073/pnas.95.3.1224.
An outbreak of human anthrax occurred in Sverdlovsk, Union of Soviet Socialists Republic (now Ekaterinburg, Russia) in April 1979. Officials attributed this to consumption of contaminated meat, but Western governments believed it resulted from inhalation of spores accidentally released from a nearby military research facility. Tissue samples from 11 victims were obtained and methods of efficiently extracting high-quality total DNA from these samples were developed. Extracted DNA was analyzed by using PCR to determine whether it contained Bacillus anthracis-specific sequences. Double PCR using "nested primers" increased sensitivity of the assay significantly. Tissue samples from 11 persons who died during the epidemic were examined. Results demonstrated that the entire complement of B. anthracis toxin and capsular antigen genes required for pathogenicity were present in tissues from each of these victims. Tissue from a vaccination site contained primarily nucleic acids from a live vaccine, although traces of genes from the infecting organisms were also present. PCR analysis using primers that detect the vrrA gene variable region on the B. anthracis chromosome demonstrated that at least four of the five known strain categories defined by this region were present in the tissue samples. Only one category is found in a single B. anthracis strain.
1979年4月，苏联社会主义共和国联盟斯维尔德洛夫斯克市（现为俄罗斯叶卡捷琳堡）爆发了人类炭疽疫情。官方将此归因于食用了受污染的肉类，但西方政府认为这是由于吸入了附近军事研究设施意外释放的孢子所致。获取了11名受害者的组织样本，并开发了从这些样本中高效提取高质量总DNA的方法。使用聚合酶链反应（PCR）对提取的DNA进行分析，以确定其是否包含炭疽芽孢杆菌特异性序列。使用“巢式引物”的双重PCR显著提高了检测的灵敏度。对疫情期间死亡的11人的组织样本进行了检查。结果表明，这些受害者每个人的组织中都存在致病性所需的完整炭疽芽孢杆菌毒素和荚膜抗原基因。接种部位的组织主要含有来自活疫苗的核酸，不过也存在感染生物体的基因痕迹。使用检测炭疽芽孢杆菌染色体上vrrA基因可变区的引物进行PCR分析表明，组织样本中存在该区域定义的五个已知菌株类别中的至少四个。在单个炭疽芽孢杆菌菌株中只发现一个类别。
