ABSTRACT
Objective To establish real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for human coronaviruses(HCoV)-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,and to investigate the prevalence of the four coronaviruses in children with acute respiratory tract infection(ARTI) in Fuzhou area.Methods Totally of 538 nasopharyngeal swabs were collected from pediatric patients with ARTI,including 289 specimens from children with acute upper respiratory tract infection (AURTI) and 249 from acute lower respiratory tract infection (AURTI) during three consecutive winter-spring seasons from December to April of 2006 - 2009 in Fuzhou area.All the specimens were subjected to FQ-PCR specific for HCoV-NL63,HCoV-HKU1,HCoV-OC43 and HCoV-229E,respectively.The enumeration data were analyzed by chi square test.Results The FQ-PCR methods were established for detecting HCoV-NL63,HCoV-HKU1,HCoV-OCA3 and HCoV-229E.The intraassay coefficient of variation (CV) and interassay CV were both ≤ 1.6％.The coronaviruses were detected in 41 (7.6％) children with ARTI,including HCoV-NL63 in 8 (1.5％)children (1 with AURTI,7 with ALRTI),HCoV-229E in 5 (0.9％; 1 with AURTI,4 with ALRTI),HCoV-HKU1 in 6 (1.1％; 1 with AURTI,5 with ALRTI),and HCoV-OC43 in 22(4.1％; 13 with AURTI,9 with ALRTI).The four coronaviruses were detected during each of the three winter-spring seasons and the positive rates of different periods were not significantly different (P＞0.05).The HCoV-OC43 positive rate was significantly higher than HCoV-NL63,HCoV-229E and HCoV-HKU1 (x2 =6.721,10.979,9.387; respectively; all P＜0.01).ConclusionsIt is suggested that the four coronaviruses might be important virus pathogens in children with ARTI in Fuzhou,China.And detection of them is needed for etiology and epidemiology evaluations for children with ARTI.