Debes Amanda K, Ateudjieu Jerome, Guenou Etienne, Ebile Walter, Sonkoua Isaac Tadzong, Njimbia Anthony Chebe, Steinwald Peter, Ram Malathi, Sack David A
Am J Trop Med Hyg. 2016 Mar;94(3):537-543. doi: 10.4269/ajtmh.15-0496. Epub 2016 Jan 11.
Biological confirmation of the presence of Vibrio cholerae in clinical and environmental samples is often constrained due to resource- and labor-intensive gold standard methods. To develop low-cost, simple, and sustainable surveillance techniques, we modified previously published specimen sampling and culture techniques and applied the use of enriched dipstick testing in conjunction with the use of filter paper for DNA specimen preservation during clinical and environmental surveillance in the Far North of Cameroon from August 2013 to October 2014. The enriched dipstick methodology during routine use in a remote setting demonstrated a specificity of 99.8% compared with polymerase chain reaction (PCR). The novel application of filter paper as a preservation method for cholera DNA specimens reduced the need for cold chain storage and allowed for PCR characterization and confirmation of V. cholerae. The application of basic technologies such as the enriched dipstick, the use of simplified gauze filtration for environmental sample collection, and the use of filter paper for sample preservation enabled early case identification with reduced logistics and supply cost while reporting minimal false-positive results. Simplified laboratory and epidemiological methodologies can improve the feasibility of cholera surveillance in rural and resource-constrained areas, facilitating early case detection and rapid response implementation.
由于资源和劳动力密集型的金标准方法,临床和环境样本中霍乱弧菌存在的生物学确认常常受到限制。为了开发低成本、简单且可持续的监测技术,我们修改了先前发表的样本采集和培养技术,并在2013年8月至2014年10月期间喀麦隆远北地区的临床和环境监测中,将富集试纸条检测与使用滤纸保存DNA样本相结合应用。在偏远地区的常规使用中,富集试纸条方法与聚合酶链反应(PCR)相比,特异性为99.8%。滤纸作为霍乱DNA样本保存方法的新应用减少了冷链储存的需求,并能够对霍乱弧菌进行PCR鉴定和确认。富集试纸条、简化纱布过滤用于环境样本采集以及滤纸用于样本保存等基础技术的应用,在减少后勤和供应成本的同时,能够以最少的假阳性结果实现早期病例识别。简化的实验室和流行病学方法可以提高农村和资源有限地区霍乱监测的可行性,促进早期病例检测和快速反应的实施。