PHE Microbiology Services, Colindale, London, UK.
Royal Free London NHS Trust, London, UK.
J Med Microbiol. 2013 Oct;62(Pt 10):1535-1539. doi: 10.1099/jmm.0.058743-0. Epub 2013 Jul 5.
A universal stool extraction method for recovery of nucleic acids (NAs) from gastrointestinal pathogens was developed to support rapid diagnostics for the London 2012 Olympics. The method involved mechanical disruption (bead beating) of the stools, followed by automated extraction and detection using real-time PCR. This method had been used extensively in the Second Infectious Intestinal Disease Study (IID2) for the isolation of NA from bacteria and parasites (and was effective for the robust recovery of Cryptosporidium spp.) but had not been used for enteric viruses. To ensure this method was universally suitable, panels of samples known to contain target bacteria, viruses or parasites were processed in triplicate using the pre-treatment method routinely used for each target and the new extraction method (bead beating). The extracts were tested using real-time PCR and the cycle threshold values were compared. The results from this study showed that bead beating improved yields for the bacterial and parasitic targets and was suitable for the viral targets. The implementation of this universal method should confer cost- and time-saving benefits and streamline the processes required for the characterization of an array of pathogens from faecal samples.
为了支持伦敦 2012 年奥运会期间的快速诊断，我们开发了一种从胃肠道病原体中提取核酸（NA）的通用粪便提取方法。该方法包括粪便的机械破坏（珠磨），然后使用实时 PCR 进行自动提取和检测。该方法已在第二次传染性肠道疾病研究（IID2）中广泛用于从细菌和寄生虫中分离 NA（并且对隐孢子虫属的有效回收），但尚未用于肠道病毒。为确保该方法具有普遍性，使用已知含有目标细菌、病毒或寄生虫的样本进行了三重复处理，使用针对每个目标的常规预处理方法和新的提取方法（珠磨）。使用实时 PCR 测试提取物，并比较循环阈值值。该研究的结果表明，珠磨法提高了细菌和寄生虫目标的产量，并且适用于病毒目标。实施这种通用方法应该可以节省成本和时间，并简化从粪便样本中对一系列病原体进行特征描述所需的过程。
