核酸序列依赖性扩增、Real-time PCR及GM试验诊断侵袭性曲霉菌感染的临床应用评价

中国真菌学杂志　 2015, Vol. 10 　Issue (5): 283-287.

核酸序列依赖性扩增、Real-time PCR及GM试验诊断侵袭性曲霉菌感染的临床应用评价

王立朋, 鲍翠霞, 于丽梅, 张晓录, 于威娟, 张霞, 李玮, 黄葆华, 李杰, 孙成铭

青岛大学医学院附属烟台毓璜顶医院检验科, 烟台 264000

收稿日期:2015-06-03 出版日期:2015-10-28 发布日期:2015-10-28
通讯作者: 黄葆华,E-mail:huangbaohua2010@126.com E-mail:huangbaohua2010@126.com
作者简介:王立朋,男(汉族),硕士,检验师.E-mail:wlp924577010@163.com

The application of nucleic acid sequence-based amplification,real-time PCR and GM test in invasive aspergillosis diagnosis

WANG Li-peng, BAO Cui-xia, YU Li-mei, ZHANG Xiao-lu, YU Wei-juan, ZHANG Xia, LI Wei, HUANG Bao-hua, LI Jie, Sun Chengming

Affiliated Yantai Yuhuangding Hospital, Medical College of Qingdao University, Yantai 264000

Received:2015-06-03 Online:2015-10-28 Published:2015-10-28

摘要/Abstract

摘要：

目的核酸序列依赖性扩增(nucleic acid sequence-based amplification,NASBA)、Real-time PCR及GM试验在侵袭性曲霉菌感染中的诊断价值。方法收集2013年11月~2014年6月临床上曲霉菌感染高危病患的血液标本80例,并根据EORTC/MSG诊断标准分为确诊组8例,拟诊组26例,非感染组46例,分别利用NASBA、real-time PCR及GM试验进行检测,计算3种方法的诊断指标并分析评价。结果 NASBA、real-time PCR及GM试验3种方法的灵敏度分别为76.47%、67.65%、52.94%,特异度分别为80.43%、89.13%、80.43%。联合诊断结果显示,NASBA与real-time PCR串联方案有最好的特异度(100%)及阳性预测值(100%);NASBA与real-time PCR并联方案则最为灵敏(94.12%)。结论 NASBA用于诊断IA最为敏感,而real-time PCR则最为特异,GM试验的表现差于前两者。联合应用的诊断策略,在特定情况下提高临床早期诊断IA的准确性。

关键词: 侵袭性曲霉菌感染, 核酸序列依赖性扩增, 实时PCR, GM试验

Abstract:

Objective To study the diagnostic performance of nucleic acid sequence-based amplification(NASBA) assay, real-time PCR and GM test in detecting invasive aspergillosis for clinical diagnosis.Methods Blood samples from 80 patients at a high risk for IA were collected during from November 2013 to June 2014.These patients were categorized as 8 proven IA, 26 probable IA, and 46 non-IA according to the 2008 revised definitions of EORTC/MSG.Blood samples were tested by NASBA, real-time PCR and GM test and their diagnostic parameters were calculated, respectively.Result The sensitivity of NASBA, real-time PCR and GM test was 76.47%, 67.65% and 52.94%, while their specificity was 80.43%, 89.13%, 80.43%, respectively.The efficiency of various combinations of tests was also evaluated.Perfect specificity(100%) and positive predictive value(100%) were achieved by combining NASBA and real-time PCR as a serial testing.A combination of NASBA and real-time PCR as a parallel testing was the most sensitive(94.12%).Conclusion The sensitivity and specificity of NASBA and real-time PCR were superior to GM test.Combination of these assays could be particularly useful in specific clinical situations.

Key words: invasive aspergillosis, nucleic acid sequence-based amplification, real-time PCR, GM test

中图分类号:

R379.6

王立朋, 鲍翠霞, 于丽梅, 张晓录, 于威娟, 张霞, 李玮, 黄葆华, 李杰, 孙成铭. 核酸序列依赖性扩增、Real-time PCR及GM试验诊断侵袭性曲霉菌感染的临床应用评价[J]. 中国真菌学杂志, 2015, 10(5): 283-287.

WANG Li-peng, BAO Cui-xia, YU Li-mei, ZHANG Xiao-lu, YU Wei-juan, ZHANG Xia, LI Wei, HUANG Bao-hua, LI Jie, Sun Chengming. The application of nucleic acid sequence-based amplification,real-time PCR and GM test in invasive aspergillosis diagnosis[J]. Chinese Journal of Mycology, 2015, 10(5): 283-287.

参考文献

[1] Chandrasekar P.Diagnostic challenges and recent advances in the early management of invasive fungalinfections[J].Eur J Haematol,2010,84(4):281-290.
[2] LeventakosK,Lewis RE,Kontoyiannis DP.Fungal infections in leukemia patients:how do we prevent and treat them[J].Clini Infect Dis,2010,50(3):405-415.
[3] Zhao Y,Park S,Kreiswirth BN,et al.Rapid real-time nucleic acid sequence based amplification-molecular beacon platform to detect fungal and bacterial bloodstream infections[J].J Clin Microbiol,2009,47(7):2067-2078.
[4] Ge Y,Cui L,Qi X,et al.Detection of novel swine origin influenza A virus(H1N1) by real-time nucleic acid sequence-based amplification[J].J Virol Methods,2010,163(2):495-497.
[5] Perlin DS,Zhao Y.Molecular diagnostic platforms for detecting Aspergillus[J].Med Mycol,2009,47(s1):S223-232.
[6] Galimberti R,Torre AC,Baztán MC,et al.Emerging systemic fungal infections[J].Clin Dermatol,2012,30(6):633-650.
[7] Dimopoulos G,Frantzeskaki F,Poulakou G,et al.Invasive aspergillosis in the intensive care unit[J].Ann N Y Acad Sci,2012,1272:31-39.
[8] De Pauw B,Walsh TJ,Donnelly JP,et al.Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group(EORTC/MSG) Consensus Group[J].Clin Infect Dis,2008,46(12):1813-1821.
[9] 高露娟,鲁巧云,孙毅,等.实时PCR在侵袭性曲霉病中的诊断价值[J].中华检验医学杂志,2013,36(2):173-177.
[10] White PL,Linton CJ,Perry MD,et al.The evolution and evaluation of a whole blood polymerase chain reaction assay for the detection of invasive aspergillosis in hematology patients in a routine clinical setting[J].Clin Infect Dis,2006,42(4):479-486.
[11] Denning DW,Kibbler CC,Barnes RA,et al.British society for medical mycology proposed standards of care for patients with invasive fungal infections[J].Lancet Infect Dis,2006,3(4):230-240.
[12] Torelli R,Sanguinetti M,Moody A,et al.Diagnosis of invasive aspergillosis by a commercial real-time PCR assay for aspergillus DNA in bronchoalveolar lavage fluid samples from high-risk patients compared to a galactomannan enzyme immunoassay[J].J Clin Microbiol,2011,49(12):4273-4278.
[13] Zhao Y,Perlin DS.Quantitative detection of Aspergillus spp. by real-time nucleic acid sequence-based amplification[J].Methods Mol Biol,2013,968:83-92.
[14] Bernal-Martínez L,Gago S,Buitrago MJ,et al.Analysis of performance of a PCR-based assay to detect DNA of Aspergillus fumigatus in whole blood and serum:a comparative study with clinical samples[J].J Clin Microbiol,2011,49(10):3596-3599.
[15] Mengoli C,Cruciani M,Barnes RA,et al.Use of PCR for diagnosis of invasive aspergillosis:systematic review and meta-analysis[J].Lancet Infect Dis,2009,9(2):89-96.
[16] Chen SC,Kontoyiannis DP.New molecular and surrogate biomarker-based tests in the diagnosis of bacterial and fungal infection in febrile neutropenic patients[J].Curr Opin Infect Dis,2010,23(6):567-577.
[17] Mennink-Kersten MA1,Ruegebrink D,Wasei N,et al.In vitro release by Aspergillus fumigatus of galactofuranose antigens,1,3-beta-D-glucan,and DNA,surrogate markers used for diagnosis of invasive aspergillosis[J].J Clin Microbiol,2006,44(5):1711-1718.