中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2013年
3期
138-143
,共6页
王璇%李莉%王睿莹%吴吉芹%胡秀平%徐斌%曹亚辉%章强强%朱利平
王璇%李莉%王睿瑩%吳吉芹%鬍秀平%徐斌%曹亞輝%章彊彊%硃利平
왕선%리리%왕예형%오길근%호수평%서빈%조아휘%장강강%주리평
真菌%RNA,核糖体%基因,真菌%序列分析,DNA
真菌%RNA,覈糖體%基因,真菌%序列分析,DNA
진균%RNA,핵당체%기인,진균%서렬분석,DNA
Fungi%RNA,ribosomal%Genes,fungal%Sequence analysis,DNA
目的 评价核糖体RNA基因(rDNA)分子鉴定用于常见致病性真菌鉴定的准确性.方法 选取致病性酵母类真菌、丝状真菌和无绿藻共60株,其中包括13株标准菌株和47株临床/环境分离株,分别对其进行表型鉴定和rDNA分子鉴定,包括内转录间隔区1(ITS1)、内转录间隔区2(ITS2)及28S亚基rDNA5'端的D1/D2区(D1/D2)靶序列的测定与比对,比较表型鉴定及单个、多个rDNA靶序列分子鉴定的准确率.结果 在23株酵母类真菌分离菌株中,D1/D2分子鉴定准确率最高(20/23,87.0％),而后依次为ITS1联合ITS2(19/23,82.6％)、单用ITS2(18/23,78.3％)、单用ITS1(14/23,60.9％)；在22株丝状真菌分离菌株中,ITS1联合ITS2分子鉴定准确率最高(16/22,72.7％),而后依次为ITS1(15/22,68.2％)、ITS2(14/22,63.6％)、D1/D2 (13/22,59.1％).而ITS1、ITS2、D1/D2三者线性联合鉴定准确率更高,可准确鉴定91.3％(21/23)的酵母类真菌分离菌株和72.7％(16/22)的丝状真菌分离菌株.结论 rDNA分子鉴定应用于常见致病性真菌的鉴定准确率高,尤其是ITS联合D1/D2具有较好的应用前景.
目的 評價覈糖體RNA基因(rDNA)分子鑒定用于常見緻病性真菌鑒定的準確性.方法 選取緻病性酵母類真菌、絲狀真菌和無綠藻共60株,其中包括13株標準菌株和47株臨床/環境分離株,分彆對其進行錶型鑒定和rDNA分子鑒定,包括內轉錄間隔區1(ITS1)、內轉錄間隔區2(ITS2)及28S亞基rDNA5'耑的D1/D2區(D1/D2)靶序列的測定與比對,比較錶型鑒定及單箇、多箇rDNA靶序列分子鑒定的準確率.結果 在23株酵母類真菌分離菌株中,D1/D2分子鑒定準確率最高(20/23,87.0％),而後依次為ITS1聯閤ITS2(19/23,82.6％)、單用ITS2(18/23,78.3％)、單用ITS1(14/23,60.9％)；在22株絲狀真菌分離菌株中,ITS1聯閤ITS2分子鑒定準確率最高(16/22,72.7％),而後依次為ITS1(15/22,68.2％)、ITS2(14/22,63.6％)、D1/D2 (13/22,59.1％).而ITS1、ITS2、D1/D2三者線性聯閤鑒定準確率更高,可準確鑒定91.3％(21/23)的酵母類真菌分離菌株和72.7％(16/22)的絲狀真菌分離菌株.結論 rDNA分子鑒定應用于常見緻病性真菌的鑒定準確率高,尤其是ITS聯閤D1/D2具有較好的應用前景.
목적 평개핵당체RNA기인(rDNA)분자감정용우상견치병성진균감정적준학성.방법 선취치병성효모류진균、사상진균화무록조공60주,기중포괄13주표준균주화47주림상/배경분리주,분별대기진행표형감정화rDNA분자감정,포괄내전록간격구1(ITS1)、내전록간격구2(ITS2)급28S아기rDNA5'단적D1/D2구(D1/D2)파서렬적측정여비대,비교표형감정급단개、다개rDNA파서렬분자감정적준학솔.결과 재23주효모류진균분리균주중,D1/D2분자감정준학솔최고(20/23,87.0％),이후의차위ITS1연합ITS2(19/23,82.6％)、단용ITS2(18/23,78.3％)、단용ITS1(14/23,60.9％)；재22주사상진균분리균주중,ITS1연합ITS2분자감정준학솔최고(16/22,72.7％),이후의차위ITS1(15/22,68.2％)、ITS2(14/22,63.6％)、D1/D2 (13/22,59.1％).이ITS1、ITS2、D1/D2삼자선성연합감정준학솔경고,가준학감정91.3％(21/23)적효모류진균분리균주화72.7％(16/22)적사상진균분리균주.결론 rDNA분자감정응용우상견치병성진균적감정준학솔고,우기시ITS연합D1/D2구유교호적응용전경.
Objective To evaluate the accuracy of ribosomal RNA gene (rDNA) molecular identification in identification of common pathogenic fungi.Methods Sixty pathogenic fungi including yeasts,filamentous fungi and Prototheca species were included in the study,among which 13 were reference strains and 47 were clinical/environmental isolates.Both phenotypic identification and rDNA molecular identification,including sequencing and alignment of internal transcribed spacer 1 (ITS1),internal transcribed spacer 2 (ITS2) and D1/D2 region on 5' end of 28S subunit rDNA (D1/D2) target regions were performed for all the 60 strains.The accuracy of single region,multiple regions molecular and phenotypic identification methods was compared.Results The highest accuracy of species-level identification for yeast isolates was yielded by D1/D2 molecular identification (20/23,87％),followed by combining ITS1 with ITS2 (19/23,82.6％),ITS2 alone(18/23,78.3％),and ITS1 alone (14/23,60.9％).While for filamentous fungal isolates,the highest species-level identification accuracy was yielded by combination of ITS1 and ITS2 molecular identification (16/22,72.7％),followed by ITS1 alone(15/22,68.2％),ITS2 alone(14/22,63.6％) and D1/D2 (13/22,59.1％).Combination of ITS1,ITS2 and D1/D2 molecular methods showed the best performance,which yielded 91.3 ％ (21/23) species-level identification for yeast isolates,and 72.7 ％ (16/22) for filamentous isolates.Conclusions rDNA molecular identification has high accuracy for identification of common pathogenic fungi,especially combination of ITS and D1/D2 regions,which might be a promising approach in the future.
