Rapid PCR of Bacillus anthracis DNA

炭疽杆菌 DNA 的快速 PCR

• 批准号: 6555512
• 负责人: NISHA V PADHYE
• 金额: $ 9.83万
• 依托单位: MEGABASE RESEARCH PRODUCTS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6555512
• 关键词: Bacillus anthracis; bacterial DNA; biotechnology; carbon dioxide; diagnosis design /evaluation; helium; polymerase chain reaction; technology /technique development

DESCRIPTION (provided by applicant): Anthrax or Woolsorter's Disease is a zoonotic infection caused by Bacillus anthracis. During the peak of the 'anthrax letter scare of 2001', the US Army Medical Research Institute for Infectious Diseases was testing over 2000 samples per day. At the present time, nearly all anthrax testing at USAMRIID relies on microbiological methods, which require one to two days to carry out. Much faster and more informative diagnosis is possible using PCR-based methods; in particular using new machines designed for high-speed amplification of DNA fragments 100 to 500 base pairs long. The fastest PCR machine employed to date is a pressurized He/CO2 gas thermocycler with digitally programmable electronic valves. Using this gas burst device, a 368 b.p. DNA fragment from B. anthracis Sterne (vaccine) strain was amplified 108-fold through 30 PCR cycles in <4 minutes. Even faster (1.5 to 3.0 minute) PCR-amplification of B. anthracis plasmid-encoded virulence factors (pag, lef, cya, capA, capC) is possible using shorter (151 -288 b.p.) amplicons and/or fewer (20-25) PCR cycles. The present experiments seek to improve the ease-of-use of this high performance He/CO2 gas thermocycler, for deployment at USAMRIID within the next year.

描述（由申请人提供）： 炭疽病或羊毛分选病是由炭疽杆菌引起的人畜共患感染。在“2001 年炭疽信件恐慌”最严重的时期，美国陆军传染病医学研究所每天检测 2000 多个样本。目前，USAMRIID 几乎所有炭疽测试都依赖微生物方法，需要一到两天才能完成。使用基于 PCR 的方法可以实现更快、信息更丰富的诊断；特别是使用专为高速扩增 100 至 500 个碱基对长的 DNA 片段而设计的新机器。迄今为止使用的最快的 PCR 机器是带有数字可编程电子阀的加压 He/CO2 气体热循环仪。使用这种气体爆破装置，可实现 368 b.p.来自炭疽芽孢杆菌 Sterne（疫苗）菌株的 DNA 片段通过 30 个 PCR 循环在不到 4 分钟内扩增了 108 倍。使用较短 (151 -288 b.p.) 扩增子和/或更少 (20-25 ) PCR 循环。目前的实验旨在提高这种高性能 He/CO2 气体热循环仪的易用性，以便于明年在 USAMRIID 进行部署。