定量PCR检测HCMV-DNA结合细胞培养分离病毒法诊断小儿HCMV感染

Diagnosis on HCMV infection in children by Real-time Fluorescence Quantitative PCR for HCMV-DNA detection and cell culture for virus isolation

目的探讨实时荧光定量PCR检测HCMV-DNA结合细胞培养分离病毒,对诊断小儿HCMV感染的价值。方法使用实时荧光定量PCR法检测尿液HCMV-DNA与细胞培养分离尿液HCMV二种方法检测。结果(1)该实时荧光定量PCR检测HCMV-DNA体系,与相关的三种病毒(I型单纯疱疹病毒、腺病毒和呼吸道合胞病毒)无交叉反应,20例正常幼儿皆为阴性。(2)40例临床患儿实时荧光定量PCR检测尿液HCMV-DNA,阳性10例。其中,唇腭裂组阳性6例(6/11),肺炎组阳性4例(4/29),两组检出结果比较,差异有显著性(χ2=5.06,P<0.05),唇腭裂组阳性检出高于肺炎组。(3)40例临床患儿细胞培养法HCMV分离检测,阳性8例,其中,唇腭裂组阳性4例(4/11),肺炎组阳性为4例(4/29),两组检出结果比较,差异无显著性(χ2=1.21,P>0.05)。(4)40例临床患儿,实时荧光定量PCR检测尿液HCMV-DNA与细胞培养分离病毒结果符合度比较,差异无显著性(χ2=3.33,P>0.05)。二种方法阳性检出比较,差异无显著性(χ2=0.38,P>0.05)。结论实时荧光定量PCR检测尿液HCMV-DNA,与细胞培养法分离病毒联合应用可提高HCMV感染诊断阳性率。

Objective To evaluate the application of Real-time Fluorescence Quantitative PCR for HCMV-DNA detection and cell culture for HCMV isolation on the diagnosis of HCMV infection in chil-dren.Methods The Real-time Fluorescence Quantitative PCR for detecting HCMV-DNA and the cell cul-ture technique for isolating HCMV in urine were used respectively,and the results were compared.Results ⑴ The Real-time Fluorescence Quantitative PCR for HCMV-DNA detection did not have cross reaction with related RSV.Adenovirus and HSV-1,and 20 normal children were all negative.⑵ 40 clinical cases were detected urine HCMV-DNA by the Real-time Fluorescence Quantitative PCR,and 10 cases were positive.6 cases were positive（6/11） in cleft lip（palate） group and 4 cases were positive（4/29） in pneumonia group.There was significant difference（χ2=5.06,P〈0.05）between the two groups.The positive rate in cleft lip（palate） group was higher than that of the pneumonia group.⑶ The 40 clinical cases were detected HCMV by vi...更多rus isolation,8 cases were positive.4 cases were positive in lip（palate） split group（4/11） and 4 cases were positive in pneumonia group（4/29）.There was no significant difference（χ2=1.21,P〉0.05） between the two groups.⑷ There was no significant difference（χ2= 3.33,P〉0.05） between the Real-time Fluorescence Quantitative PCR and the cell culture technique on the diagnosis of HCMV infection in children.Conclusion The Real-time Fluorescence Quantitative PCR for detecting HCMV-DNA combined with the cell culture technique for iso-lating HCMV in urine can raise the positive rate on the diagnosis of HCMV infection in children. Objective To compare the methods of ATBFUNGUS2 microdilution,the ROSCO disk diffusion and the NCCLS disk diffusion in extracorporeal antifungal susceptibility testing.Methods A total of 105 clinical fungal isolates were tested for susceptibility to fluconazole,itraconazole,AMB,5-FC by the three methods respectively.Results The results of the antifungal susceptibility testing of the three methods had fine consistency and repeatability.Conclusion The methods of ATBFUNGUS2 microdilution and the Rosco disk diffusion can be used as extracorporeal antifungal susceptibility testing in clinic laboratories.