返回Google圖書搜尋

Validation Report of a Real-time PCR Assay for the Identification of Clavibacter Michiganensis Subsp. Michiganensis (Cmm) Isolates

出版	Groen Agro Control, 2010
URL	http://books.google.com.hk/books?id=kX4WogEACAAJ&hl=&source=gbs_api

註釋This reports presents complete validation data related to recently developed primer-probe combinations for the identification of Clavibacter michiganensis subsp. michiganensis (Cmm) isolates with the use of a real-time PCR assay. The primers and probes were developed on two loci: on the intergenic spacer between the 16S and 23S ribosomal RNA genes (IGS) and on the pathogenity-1 gene (Pat-1), which is located on the plasmid pCM2. The validated method is qualitative and consists of a DNA extraction from pure colony material followed by a duplex real-time PCR assay based on the primer-probe combinations for IGS and Pat-1. The method is very specific, because only the 20 tested pathogenic Cmm isolates were positive for both primer-probe combinations. Negative results were obtained from 12 Cmm look-alikes, 4 Cm isolates and 9 other tomato pathogens. Tests on repeatability, reproducibility and robustness of this assay performed well.