Liu D, Pearce L, Lilley G, Coloe S, Baird R, Pedersen J
Melbourne Pathology, Collingwood, Victoria, Australia.
Med Mycol. 2001 Apr;39(2):215-9. doi: 10.1080/mmy.39.2.215.219.
A DNA fragment of approximately 1.2 kb, generated from the common dermatophyte Microsporum canis by arbitrarily primed polymerase chain reaction (PCR) using random primer OPU13, was cloned and sequenced. Based on the resulting sequencing data, a forward primer (MC1F) and a reverse primer (MC1R) have been designed and assessed by PCR for their usefulness in the improved identification of M. canis. The results obtained suggest that these primers are specific for M. canis, as a band of 900 bp was amplified in PCR with genomic DNA from M. canis only, and not from any of the other dermatophyte species or varieties, other fungi or common bacteria examined. Combining this PCR technique with a rapid mini-preparation method for fungal DNA, a definitive diagnosis of M. canis can be achieved within a day from the primary cultures. Future refinement of a DNA purification protocol from clinical specimens would further enhance the potential of the PCR based test for improved detection and identification of M. canis.
使用随机引物OPU13通过任意引物聚合酶链反应（PCR）从常见皮肤癣菌犬小孢子菌中产生的一段约1.2 kb的DNA片段被克隆并测序。基于所得的测序数据，设计了正向引物（MC1F）和反向引物（MC1R），并通过PCR评估了它们在改进犬小孢子菌鉴定中的实用性。获得的结果表明，这些引物对犬小孢子菌具有特异性，因为在仅用犬小孢子菌的基因组DNA进行的PCR中扩增出了一条900 bp的条带，而在所检测的其他皮肤癣菌物种或变种、其他真菌或常见细菌中均未扩增出该条带。将这种PCR技术与一种快速的真菌DNA微量制备方法相结合，从原代培养物开始，一天内即可实现对犬小孢子菌的确切诊断。临床标本DNA纯化方案的进一步完善将进一步提高基于PCR的检测方法在改进犬小孢子菌检测和鉴定方面的潜力。
