Klinger J D, Johnson A, Croan D, Flynn P, Whippie K, Kimball M, Lawrie J, Curiale M
GENE-TRAK Systems, Framingham, MA 01701.
J Assoc Off Anal Chem. 1988 May-Jun;71(3):669-73.
A nucleic acid hybridization assay has been developed for Listeria spp. in dairy foods and environmental samples. The assay is based on detection of unique Listeria 16S rRNA sequences by using a 32P-labeled synthetic DNA probe. Inclusivity and exclusivity of the probe were confirmed with 139 Listeria isolates representing all known species, and 73 non-Listeria bacterial strains. In this paper, we present results from our preliminary studies comparing the hybridization assay with conventional culture on a total of 575 specimens that represent a variety of inoculated and uninoculated foods and environmental samples. The assay, which is done in a filter manifold format after 2 days of cultural enrichment, requires a total assay time of less than 2.5 days. The false-negative rate for all sample groups tested using the GENE-TRAK hybridization assay was less than the rate for culture. Thus, the new assay allows rapid screening of the indicated product groups and provides reliable numerical results.
已开发出一种用于检测乳制品和环境样品中李斯特菌属的核酸杂交检测方法。该检测方法基于使用32P标记的合成DNA探针检测独特的李斯特菌16S rRNA序列。用代表所有已知物种的139株李斯特菌分离株和73株非李斯特菌菌株确认了探针的包容性和排他性。在本文中,我们展示了初步研究的结果,该研究将杂交检测方法与传统培养方法在总共575个标本上进行了比较,这些标本代表了各种接种和未接种的食品及环境样品。该检测方法在经过2天的培养富集后以滤膜歧管形式进行,总检测时间不到2.5天。使用GENE-TRAK杂交检测方法检测的所有样品组的假阴性率低于培养法的假阴性率。因此,新的检测方法能够对指定的产品组进行快速筛选,并提供可靠的数值结果。