The Appraisement Of Diagnosising HCMV Production Infection By Real-time Fluorescence Quantitative PCR For The Detection Of HCMV-DNA In Children

Objective To establish the Real-time Fluorescence Quantitative PCR for the detection of HCMV-DNA and to appraise its application value of the method in children HCMV infection diagonsis.Methods Establishing the Real-time Fluorescence Quantitative PCR and evaluating the credibility of this method by analysising the results of three HCMV relative virus strains and 20 infants urine samples as control.Collecting the blood and urine samples of children including 29 cases of pneumonia and 11 cases of cleft lip(palate),Three methods were applied:1.The Real-time Fluorescence Quantitative PCR for detecting HCMV-DNA;2.The cell culture for segregating HCMV;3.ELISA for detectiong HCMV-IgG and HCMV-IgM.Meanwhile,the white blood cell count and urine protein qualitative analysis are tested in these 40 cases.Results:1.The Real-time Fluorescence Quantitative PCR system for the detection of HCMV-DNA,with related RSV Adenovirus HSV do not have overlapping reaction, 20 normal children are negative(＜100 copies/m1 ).2.40 clinical cases were detected urine HCMV-DNA by the Real-time Fluorescence Quantitative PCR,the positive is 10 cases.The positive of cleft lip (palate) group is 6 cases(6/11 ),The positive of pneumonia group is 4 cases (4/29 ).The rate of positive is significance difference(X~2=5.06,P＜0.05) between the two groups.The rate of positive in cleft lip(palate) group is higher than in pneumonia group.3.40 clinical cases were detected blood plasma HCMV-IgM by ELISA,the positive is 8 cases.The positive of cleft lip(palate) group is 6 cases(6/11 ), The positive of pneumonia group is 2 cases(2/29 ).The rate of positive is significance difference(X~2=8.53,P＜0.05) between the two groups,The rate of positive in cleft lip(palate) group is higher than in pneumonia group.4.40 clinical cases were detected the HCMV cell by culture method.The positive is 8 cases.The positive of lip palate split group is 4 cases(4/11 ),The positive of pneumonia group is 4 cases(4/29 ).The rate of positive is not significance difference(X~2=1.21,P＞0.05) between the two groups.5.In 40 clinical cases the result of Real-time Fluorescence Quantitative PCR and the HCMV cell by culture method have not significance difference (X~2= 3.33,P＞0.05 ),ELISA detection HCMV-IgM of blood plasma and the Real-time Fluorescence Quantitative PCR result have significance difference(X~2= 7.5,P＜0.05).ELISA detection HCMV-IgM of blood plasma and the HCMV cell by culture method have significance difference(X~2= 11.29,P＜0.05 ).6.In 40 clinical cases white blood cell count,do not have significance difference between urine HCMV-DNA positive group and negative group.7.40 clinical cases urine protein determine the results for negative.Conclusion:The Real-time Fluorescence Quantitative PCR detecting urine HCMV-DNA,for diagnosing HCMV production infection have certain suitable clinical value;This method combination ELISA detection serum IgM antibody can raise the detection rate of HCMV production infection,and can judge HCMV primary infection,recurrent infection or latent infection...