通过聚合酶链反应检测火鸡支原体。

Detection of Mycoplasma meleagridis by polymerase chain reaction.

作者信息

Zhao S, Yamamoto R

机构信息

Department of Epidemiology and Preventive Medicine, School of Veterinary Medicine, University of California, Davis 95616.

出版信息

Vet Microbiol. 1993 Jul;36(1-2):91-7. doi: 10.1016/0378-1135(93)90131-p.

DOI:10.1016/0378-1135(93)90131-p

PMID:8236783

Abstract

A pair of 32 base primers was synthesized based on the DNA sequence data of a Mycoplasma meleagridis (MM) species-specific recombinant, pMM-2. The primers were used in a MM-polymerase chain reaction (PCR) to amplify a target DNA of approximately 850 bp. Annealing temperatures ranging from 58 degrees C to 61 degrees C could be used for the MM-PCR without loss of specificity. The primers amplified 1 ng of DNA from 17 strains of MM, but not 10 ng of DNA from 16 heterologous species of avian mycoplasma, pUC8 plasmid, lambda phage or calf thymus DNA. The minimum amount of target DNA detected by MM-PCR was 10 fg, which indicated that this procedure was 100000 times more sensitive than dot blot methodology using an MM recombinant DNA probe.

摘要

根据火鸡支原体（MM）种特异性重组体pMM - 2的DNA序列数据合成了一对32个碱基的引物。这些引物用于MM聚合酶链反应（PCR），以扩增约850 bp的目标DNA。58℃至61℃的退火温度可用于MM - PCR，且不会丧失特异性。这些引物能从17株MM中扩增出1 ng DNA，但不能从16种禽支原体异源物种、pUC8质粒、λ噬菌体或小牛胸腺DNA的10 ng DNA中扩增出产物。MM - PCR检测到的目标DNA的最小量为10 fg，这表明该方法比使用MM重组DNA探针的斑点印迹法灵敏100000倍。