国际检验医学杂志
國際檢驗醫學雜誌
국제검험의학잡지
INTERNATIONAL JOURNAL OF LABORATORY MEDICINE
2014年
12期
1580-1582
,共3页
穿刺引流液%病原性真菌%内部转录间隔区%测序%快速检测
穿刺引流液%病原性真菌%內部轉錄間隔區%測序%快速檢測
천자인류액%병원성진균%내부전록간격구%측서%쾌속검측
puncture fluid%pathogenic fungi%internal transcribed spacer%sequencing%rapid detection
目的：探讨内部转录间隔区（ITS）测序用于快速检测穿刺引流液中病原性真菌的价值。方法采用临床常见的14种细菌和4种真菌及人类基因组DNA验证真菌通用引物（ITS1/ITS4和ITS3/ITS4）的特异性和敏感性；采集90例临床疑似真菌感染患者的穿刺引流液标本10 mL，其中8 mL 用于常规真菌培养，剩余2 mL 用于提取真菌 DNA 并分别用 ITS1/ITS4和ITS3/ITS4作为引物进行扩增，将阳性扩增产物测序并与BLAST比对，将测序法与培养法的阳性率结果进行统计学比较。结果14种细菌及人类基因组扩增产物均为阴性，4种真菌扩增产物为阳性。ITS3/ITS4和 ITS1/ITS4的最低检测限分别为102 CFU/mL和103 CFU/mL。90例标本培养阳性率为4．44％（4/90），测序法阳性率为12．22％（11/90），二者差异具有统计学意义（P＜0．05）。结论引物ITS1/ITS4和ITS3/ITS4的特异性良好，后者较前者扩增敏感性更高；测序真菌ITS区可作为临床快速检测穿刺引流液中病原性真菌感染的方法。
目的：探討內部轉錄間隔區（ITS）測序用于快速檢測穿刺引流液中病原性真菌的價值。方法採用臨床常見的14種細菌和4種真菌及人類基因組DNA驗證真菌通用引物（ITS1/ITS4和ITS3/ITS4）的特異性和敏感性；採集90例臨床疑似真菌感染患者的穿刺引流液標本10 mL，其中8 mL 用于常規真菌培養，剩餘2 mL 用于提取真菌 DNA 併分彆用 ITS1/ITS4和ITS3/ITS4作為引物進行擴增，將暘性擴增產物測序併與BLAST比對，將測序法與培養法的暘性率結果進行統計學比較。結果14種細菌及人類基因組擴增產物均為陰性，4種真菌擴增產物為暘性。ITS3/ITS4和 ITS1/ITS4的最低檢測限分彆為102 CFU/mL和103 CFU/mL。90例標本培養暘性率為4．44％（4/90），測序法暘性率為12．22％（11/90），二者差異具有統計學意義（P＜0．05）。結論引物ITS1/ITS4和ITS3/ITS4的特異性良好，後者較前者擴增敏感性更高；測序真菌ITS區可作為臨床快速檢測穿刺引流液中病原性真菌感染的方法。
목적：탐토내부전록간격구（ITS）측서용우쾌속검측천자인류액중병원성진균적개치。방법채용림상상견적14충세균화4충진균급인류기인조DNA험증진균통용인물（ITS1/ITS4화ITS3/ITS4）적특이성화민감성；채집90례림상의사진균감염환자적천자인류액표본10 mL，기중8 mL 용우상규진균배양，잉여2 mL 용우제취진균 DNA 병분별용 ITS1/ITS4화ITS3/ITS4작위인물진행확증，장양성확증산물측서병여BLAST비대，장측서법여배양법적양성솔결과진행통계학비교。결과14충세균급인류기인조확증산물균위음성，4충진균확증산물위양성。ITS3/ITS4화 ITS1/ITS4적최저검측한분별위102 CFU/mL화103 CFU/mL。90례표본배양양성솔위4．44％（4/90），측서법양성솔위12．22％（11/90），이자차이구유통계학의의（P＜0．05）。결론인물ITS1/ITS4화ITS3/ITS4적특이성량호，후자교전자확증민감성경고；측서진균ITS구가작위림상쾌속검측천자인류액중병원성진균감염적방법。
Objective To explore the value of sequencing internal transcribed spacer(ITS)in identification of pathogenic fungi species from the puncture fluid.Methods The specificities and sensitivities of primers(ITS1/ITS4 and ITS3/ITS4)were validated by using 14 kinds of bacteria,4 kinds of fungi and human DNA.90 cases of clinical patients with suspected fungal infections were enrolled.10 mL puncture drainage fluid was collected from each patient,and 8 mL in which was cultured and the rest 2 mL was used for DNA extraction and PCR detection with ITS1/ITS4 and ITS3/ITS4 as primers.The positive PCR products were compared with BLAST,and the results were analyzed statistically.Results PCR products of bacteria and human DNA were negative,which of fungi were positive.The lowest detectable limits of ITS1/ITS4 and ITS3/ITS4 were 103 CFU/mL and 102 CFU/mL,respective-ly.Of the 90 cases of puncture drainage fluid samples,the cultivation positive rate was 4.44%(4/90),PCR positive rate was 12.22%(11/90),and the difference was statistically significant(P<0.05).Conclusion ITS1/ITS4 and ITS3/ITS4 are well in spe-cificity,but sensitivity of ITS3/ITS4 is higher than ITS1/ITS4 .ITS region sequencing will become a quick method of detecting fun-gal infection in the puncture fluid.
