基于分子检测方法在可重复灌装饮用水样品中的敏感性和特异性

A. Achyar, I. Putri, D. Putri
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摘要

可续水饮水站的存在帮助社区获得负担得起和实用的饮用水。然而,劣质饮用水会对健康产生影响。适于饮用的饮用水质量参数之一是不受大肠杆菌、沙门氏菌、埃希氏菌等细菌污染。饮用水质量的测定,除微生物学检测外,还可采用分子PCR (Polymerase Chain Reaction,聚合酶链反应)法进行。因此,本研究的目的是检验PCR检测饮用水病原体的敏感性和特异性。从大肠杆菌、沙门氏菌、埃希氏菌和一些非大肠菌群细菌培养物中提取DNA。分别用大肠杆菌- aa - forward和大肠杆菌- aa - reverse、沙门氏菌- y - forward和沙门氏菌- y - reverse、大肠杆菌- db - forward和大肠杆菌- db - reverse引物对进行PCR。结果表明,该方法对大肠杆菌、沙门氏菌、埃希氏菌的最低DNA检出量分别为0.0025 ng/µL、0,0005 ng/µL、0,04 ng/µL。各引物对的PCR特异性结果表明,该方法对大肠杆菌、沙门氏菌和埃希氏菌的PCR产物大小分别为±417 bp、±559 bp和±815 bp,均能特异性检测出大肠杆菌。
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Sensitivity and Specificity of Molecular-Based Pathogen Detection Method in Refillable Drinking Water Samples
The existence of refillable drinking water depots helps the community to get affordable and practical drinking water. However, poor quality drinking water will, however, have an effect on health. One of the quality parameters of drinking water that is suitable for consumption is not contaminated by the bacteria Escherichia coli, Salmonella sp. and Escherichia sp. Measurement of the quality of drinking water, in addition to microbiological tests, can be carried out molecularly using PCR (Polymerase Chain Reaction) method. Therefore, the aim of this study was to examine the sensitivity and specificity of PCR for detection of drinking water pathogens. DNA was extracted from cultures of E. coli, Salmonella sp., Escherichia sp. and some non-coliform bacteria. PCR was performed separately using primer pairs of E. coli-AA-Forward and E.coli-AA-Reverse, Salmonella-OY-Forward and Salmonella-OY-Reverse, E. coli-DB-Forward and E. coli-DB-Reverse. The results of the PCR sensitivity showed that the minimum amount of DNA that can be detected by this method were 0.0025 ng/µL, 0,0005 ng/µL, 0,04 ng/µL for E. coli, Salmonella sp., Escherichia sp., respectively. The results of the PCR specificity of each primer pairs indicated that these methods were able to detect each coliform bacterium specifically according to PCR product size of ± 417 bp, ± 559 bp and ± 815 bp for E. coli, Salmonella sp., Escherichia sp., respectively.
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