Department of Medical Microbiology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
PLoS One. 2011;6(10):e24830. doi: 10.1371/journal.pone.0024830. Epub 2011 Oct 3.
We describe a test for rapid detection of S. dysenteriae 1 in bacterial cultures and in stools, at the bedside of patients.
METHODOLOGY/PRINCIPAL FINDINGS: The test is based on the detection of S. dysenteriae 1 lipopolysaccharide (LPS) using serotype 1-specific monoclonal antibodies coupled to gold particles and displayed on a one-step immunochromatographic dipstick. A concentration as low as 15 ng/ml of LPS was detected in distilled water and in reconstituted stools in 10 minutes. In distilled water and in reconstituted stools, an unequivocal positive reaction was obtained with 1.6×10⁶ CFU/ml and 4.9×10⁶ CFU/ml of S. dysenteriae 1, respectively. Optimal conditions to read the test have been determined to limit the risk of ambiguous results due to appearance of a faint yellow test band in some negative samples. The specificity was 100% when tested with a battery of Shigella and unrelated strains in culture. When tested on 328 clinical samples in India, Vietnam, Senegal and France by laboratory technicians and in Democratic Republic of Congo by a field technician, the specificity (312/316) was 98.7% (95% CI:96.6-99.6%) and the sensitivity (11/12) was 91.7% (95% CI:59.8-99.6%). Stool cultures and the immunochromatographic test showed concordant results in 98.4 % of cases (323/328) in comparative studies. Positive and negative predictive values were 73.3% (95% CI:44.8-91.1%) and 99.7% (95% CI:98-100%).
The initial findings presented here for a simple dipstick-based test to diagnose S. dysenteriae 1 demonstrates its promising potential to become a powerful tool for case management and epidemiological surveys.
我们描述了一种用于快速检测患者床边细菌培养物和粪便中志贺氏菌 1 的检测方法。
方法/主要发现:该检测方法基于使用与金颗粒偶联的血清型 1 特异性单克隆抗体检测志贺氏菌 1 脂多糖(LPS),并显示在一步免疫层析试纸上。在蒸馏水中和重建的粪便中,该检测方法可在 10 分钟内检测到低至 15ng/ml 的 LPS。在蒸馏水中和重建的粪便中,分别用 1.6×10⁶ CFU/ml 和 4.9×10⁶ CFU/ml 的志贺氏菌 1 得到明确的阳性反应。为了限制由于某些阴性样本中出现淡黄色测试带而导致结果不确定的风险,已经确定了最佳的测试条件。当用培养的一系列志贺氏菌和无关菌株进行测试时,特异性为 100%。在印度、越南、塞内加尔和法国由实验室技术人员以及在刚果民主共和国由现场技术人员对 328 份临床样本进行测试时,特异性(312/316)为 98.7%(95%CI:96.6-99.6%),敏感性(11/12)为 91.7%(95%CI:59.8-99.6%)。在比较研究中,粪便培养物和免疫层析检测在 98.4%的病例(323/328)中显示出一致的结果。阳性和阴性预测值分别为 73.3%(95%CI:44.8-91.1%)和 99.7%(95%CI:98-100%)。
本文初步研究了一种简单的基于试条的检测方法来诊断志贺氏菌 1,证明其具有成为病例管理和流行病学调查的有力工具的潜力。