@article{mbs:/content/journal/jmm/10.1099/jmm.0.000266, author = "Chen, Yi and He, Hui and Pan, Ping and He, Songzhe and Dong, Xueyan and Chen, Yueming and Wang, Shuying and Yu, Daojun", title = "Rapid and combined detection of Mycoplasma pneumoniae, Epstein–Barr virus and human cytomegalovirus using AllGlo quadruplex quantitative PCR", journal= "Journal of Medical Microbiology", year = "2016", volume = "65", number = "7", pages = "590-595", doi = "https://doi.org/10.1099/jmm.0.000266", url = "https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.000266", publisher = "Microbiology Society", issn = "1473-5644", type = "Journal Article", keywords = "Mycoplasma pneumoniae", keywords = "internal control", keywords = "AllGlo probes", keywords = "Epstein–Barr virus", keywords = "quadruplex quantitative PCR", 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 (r 2) 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.", }