Rapid and combined detection of Mycoplasma pneumoniae, Epstein-Barr virus and human cytomegalovirus using AllGlo quadruplex quantitative PCR.
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.
Affiliated First Hospital of Hangzhou, Zhejiang Chinese Medical University, Hangzhou, PR China.|Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Yi
Y
Chen
Affiliated First Hospital of Hangzhou, Zhejiang Chinese Medical University, Hangzhou, PR China.|Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Hui
H
He
Affiliated First Hospital of Hangzhou, Zhejiang Chinese Medical University, Hangzhou, PR China.|Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Ping
P
Pan
Department of Clinical Laboratory, Affiliated Hospital of Guilin Medical University, Guilin, PR China.
Songzhe
S
He
Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Xueyan
X
Dong
Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Yueming
Y
Chen
Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Shuying
S
Wang
Affiliated First Hospital of Hangzhou, Zhejiang Chinese Medical University, Hangzhou, PR China.|Department of Clinical Laboratory, Hangzhou First People's Hospital, Hangzhou, PR China.
Daojun
D
Yu
D060085:Coinfection / Q000175:diagnosis; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis; D020031:Epstein-Barr Virus Infections / Q000175:diagnosis; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009175:Mycoplasma Infections / Q000175:diagnosis; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis*; D013997:Time Factors