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禽饲料中沙门氏菌检测的培养方法与聚合酶链反应检测法的比较研究

A comparative study of culture methods and polymerase chain reaction assay for Salmonella detection in poultry feed.

机构信息

Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria, Concepción del Uruguay, Entre Ríos, Argentina.

出版信息

Poult Sci. 2011 Nov;90(11):2606-18. doi: 10.3382/ps.2011-01548.

Abstract

The present work compared 2 culture methods and PCR assay for the detection of motile and non-motile Salmonella strains using artificially contaminated poultry feed. The specificity was 1 in all methods. The accuracy and sensitivity were between 0.5 and 1 for motile Salmonella strains, whereas these parameters were between 0 and 0.6 for non-motile Salmonella strains. The positive predictive value was 1 for tetrathionate (TT), PCR, and modified semisolid Rappaport-Vassiliadis (MSRV) methods in most of the strains studied. The negative predictive value of each method was very low for non-motile Salmonella strains. The detection level of motile strains was 8 to 20 cfu/25 g for all methods, whereas it was ≥10(4) cfu/25 g in culture methods for non-motile Salmonella strains. In general, the PCR method detected lower non-motile Salmonella contamination levels in feed than did culture methods. Extending incubation time of the enrichment medium to 6 d in the TT method did not improve the isolation rates. All selective plating media did not show any statistical differences in the parameters of performance studied. Kappa coefficients showed that there was good agreement between TT and MSRV methods, and MSRV and PCR methods for motile Salmonella strains in poultry feed samples. The agreement was fair between TT and PCR methods for these strains. For non-motile Salmonella strains, there was poor (TT and MSRV methods), slight (PCR and TT methods), and fair (MSRV and PCR methods) agreement. The TT, MSRV, and PCR methods are similar in terms of accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for different motile Salmonella strains in poultry feed. For non-motile Salmonella strains, the use of the PCR method improves the same parameters, described before, in this matrix. The difference in detection levels obtained with the methods used for motile and nonmotile Salmonella strains and the difficulty to detect these last strains represent a potential problem, when a poultry feed sample is considered negative for the presence of Salmonella.

摘要

本研究比较了 2 种培养方法和聚合酶链反应(PCR)检测法,用于检测人工污染的家禽饲料中运动性和非运动性沙门氏菌菌株。所有方法的特异性均为 1。对于运动性沙门氏菌菌株,准确性和敏感性在 0.5 到 1 之间,而对于非运动性沙门氏菌菌株,这些参数在 0 到 0.6 之间。在大多数研究的菌株中,四硫磺酸钠(TT)、PCR 和改良半固体雷帕氏(MSRV)方法的阳性预测值均为 1。对于非运动性沙门氏菌菌株,每种方法的阴性预测值均非常低。所有方法对运动性菌株的检测水平为 25g 饲料中 8 到 20cfu,而对于非运动性沙门氏菌菌株,培养方法的检测水平为≥10(4)cfu/25g。一般来说,PCR 方法在饲料中检测到的非运动性沙门氏菌污染水平低于培养方法。将 TT 法的增菌时间延长至 6d 并未提高分离率。所有选择性平板培养基在研究的性能参数方面均无统计学差异。Kappa 系数表明,TT 法和 MSRV 法、MSRV 法和 PCR 法之间对禽用饲料中运动性沙门氏菌菌株的检测结果具有良好的一致性。对于这些菌株,TT 法和 PCR 法之间的一致性为中等。对于非运动性沙门氏菌菌株,一致性较差(TT 法和 MSRV 法)、轻微(PCR 法和 TT 法)和中等(MSRV 法和 PCR 法)。对于禽用饲料中的不同运动性沙门氏菌菌株,TT、MSRV 和 PCR 方法在准确性、敏感性、特异性、阳性预测值和阴性预测值方面具有相似性。对于非运动性沙门氏菌菌株,PCR 方法提高了该基质中上述参数的检测能力。运动性和非运动性沙门氏菌菌株的检测水平差异以及难以检测这些最后菌株,代表了在考虑禽用饲料样品中不存在沙门氏菌时可能存在的问题。

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