CAJ | 학술논문

目的探讨菌落PCR在检测病原性丝状真菌方面的应用价值.方法初步建立用于丝状真菌的菌落PCR检测技术,用19种丝状真菌标准株进行验证,所有菌落PCR扩增产物进行测序,并选取8种菌株的菌落PCR产物和酶切结果与常规PCR进行比较,检测其准确性和可靠性.结果 19株菌中有16株(84.2%)菌落PCR成功扩增内转录间隔(ITS)区,ITS区基因序列分析鉴定菌种正确,与NCBI数据库中相同菌种的相似度为96%～100%;与常规PCR进行比较的8株菌中,除构巢曲霉菌落PCR扩增结果为阴性外,其他菌种菌落PCR产物及酶切条带与常规PCR基本一致.结论与常规PCR相比,菌落PCR检测丝状真菌操作简单,省时省力,鉴定菌种具有较高的准确性和可靠性,可以用于丝状真菌的快速鉴定.
목적 탐토균락PCR재검측병원성사상진균방면적응용개치.방법 초보건립용우사상진균적균락PCR검측기술,용19충사상진균표준주진행험증,소유균락PCR확증산물진행측서,병선취8충균주적균락PCR산물화매절결과여상규PCR진행비교,검측기준학성화가고성.결과 19주균중유16주(84.2%)균락PCR성공확증내전록간격(ITS)구,ITS구기인서렬분석감정균충정학,여NCBI수거고중상동균충적상사도위96%～100%;여상규PCR진행비교적8주균중,제구소곡매균락PCR확증결과위음성외,기타균충균락PCR산물급매절조대여상규PCR기본일치.결론 여상규PCR상비,균락PCR검측사상진균조작간단,성시성력,감정균충구유교고적준학성화가고성,가이용우사상진균적쾌속감정.
Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.