Wolf Sandro, Hewitt Joanne, Rivera-Aban Malet, Greening Gail E
Communicable Disease Group, Institute of Environmental Science & Research Ltd., Kenepuru Science Centre, PO Box 50-348, Porirua, New Zealand.
J Virol Methods. 2008 Apr;149(1):123-8. doi: 10.1016/j.jviromet.2007.12.012. Epub 2008 Feb 15.
Genotyping of F+ RNA bacteriophages has been used to distinguish between human and animal contributions to contaminated water and food. There are four genetically distinct genogroups of F+ RNA bacteriophages. Genogroups I and IV predominate in animal wastes and genogroups II and III in wastes of human origin. In this study, a multiplex real-time RT-PCR-based method was developed to detect and genotype F+ RNA bacteriophages. The assay was shown to be broadly reactive against a wide spectrum of F+ RNA bacteriophage strains, including MS2, GA, Q beta, MX1, SP and FI, and was able to detect and genotype F+ RNA bacteriophages in shellfish and river water. The assay is highly sensitive, with detection limits <10 PFU/reaction and <10 copies/reaction of the target sequences carried in plasmids, respectively. The applications of this assay include F+ RNA semi-quantitation and microbial source tracking.
F+ RNA噬菌体的基因分型已被用于区分人类和动物对受污染水和食物的影响。F+ RNA噬菌体有四个基因不同的基因组。基因组I和IV在动物粪便中占主导地位，而基因组II和III在人类来源的粪便中占主导地位。在本研究中，开发了一种基于多重实时RT-PCR的方法来检测F+ RNA噬菌体并进行基因分型。该检测方法对包括MS2、GA、Qβ、MX1、SP和FI在内的多种F+ RNA噬菌体菌株具有广泛的反应性，并且能够检测贝类和河水中的F+ RNA噬菌体并进行基因分型。该检测方法高度灵敏，检测限分别为<10 PFU/反应和<10拷贝/反应的质粒携带的靶序列。该检测方法的应用包括F+ RNA的半定量和微生物源追踪。
