Wang Haiyan
Health Canada, Bureau of Microbial Hazards, Health Products and Food Branch, Food Directorate, Ottawa, Ontario.
J AOAC Int. 2002 Jul-Aug;85(4):996-9.
Campylobacter spp. are the most commonly reported bacterial cause of acute diarrheal disease in humans throughout the world. Traditional cultural methods for the detection and quantitation of Campylobacterspp. are slow and tedious; therefore, specific, sensitive, and rapid methods for campylobacters are needed to collect sufficient data for risk assessment and food safety policy development. We developed several rapid methods based on polymerase chain reaction (PCR), DNA hybridization, hydrophobic grid membrane filters (HGMFs), and enzyme immunoassays (EIAs). A PCR assay targeting C. jejuni, combined with a simple sample preparation procedure, detects as few as 0.3 most probable number (MPN)/mL C. jejuni in naturally contaminated chicken rinses after 20-24 h enrichment. An HGMF-EIA method using a commercial polyclonal antibody for Campylobacter detects and enumerates thermophilic Campylobacter spp. from spiked chicken rinse and milk, and naturally contaminated chicken rinses. A C. jejuni-specific probe in an HGMF-DNA hybridization protocol specifically detects and quantitates C. jejuni in food samples. A dot-blot EIA combined with an MPN procedure quantitates thermophilic campylobacters from samples that might be difficult to filter through HGMFs.
弯曲杆菌属是全世界人类急性腹泻疾病中最常报告的细菌性病因。用于检测和定量弯曲杆菌属的传统培养方法缓慢且繁琐；因此，需要针对弯曲杆菌的特异性、灵敏且快速的方法，以收集足够的数据用于风险评估和食品安全政策制定。我们基于聚合酶链反应（PCR）、DNA杂交、疏水网格膜过滤器（HGMF）和酶免疫测定（EIA）开发了几种快速方法。一种针对空肠弯曲菌的PCR检测方法，结合简单的样品制备程序，在富集20 - 24小时后，可检测出自然污染鸡肉冲洗液中低至0.3最可能数（MPN）/mL的空肠弯曲菌。一种使用针对弯曲杆菌的商业多克隆抗体的HGMF - EIA方法，可从加标的鸡肉冲洗液和牛奶以及自然污染的鸡肉冲洗液中检测和计数嗜热弯曲杆菌属。HGMF - DNA杂交方案中的空肠弯曲菌特异性探针可特异性检测和定量食品样品中的空肠弯曲菌。一种与MPN程序结合的斑点印迹EIA可对可能难以通过HGMF过滤的样品中的嗜热弯曲杆菌进行定量。
