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J Med Microbiol. 2016 Jul;65(7):590-595. doi: 10.1099/jmm.0.000266. Epub 2016 Apr 19.

Rapid and combined detection of Mycoplasma pneumoniae, Epstein-Barr virus and human cytomegalovirus using AllGlo quadruplex quantitative PCR.

Chen Y1,2, He H1,2, Pan P1,2, He S3, Dong X2, Chen Y2, Wang S2, Yu D1,2.

Author information

1
1​Affiliated First Hospital of Hangzhou, Zhejiang Chinese Medical University, Hangzhou, PR China.
2
2​Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
3
3​Department of Clinical Laboratory, Affiliated Hospital of Guilin Medical University, Guilin, PR China.

Abstract

Acute respiratory infections (ARIs) cause substantial morbidity and mortality worldwide. The causes of ARI are dynamic, and co-infections of Mycoplasma pneumoniae, Epstein-Barr virus and human cytomegalovirus are recently developed causes of ARI. Here, we established a quadruplex quantitative PCR (qPCR) method to rapidly identify and simultaneously detect a single infection or co-infection of these three pathogens and an internal control in a single tube using AllGlo probes. The analysis demonstrated a wide linear range of detection from 101 to 108 copies per test and a low coefficient of variation of less than 5 %. The amplification efficiencies were all close to 1, and the correlation coefficients (r2) were all greater than 0.99. We found no significant difference in a comparative reagent test (P >0.05). Moreover, the results of tests on clinical samples using AllGlo quadruplex qPCR and TaqMan uniplex qPCR were in near-perfect agreement (κ =0.97). Clinically, the availability of this method will enable better differential diagnosis, disease surveillance and controlled outcomes.

PMID:
27093597
DOI:
10.1099/jmm.0.000266
[Indexed for MEDLINE]

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