福州地区急性呼吸道感染儿童人冠状病毒NL63、229E、HKU1和OC43的检测与分析

Detection of human coronaviruses NL63, 229E,HKU1 and OC43 in children with acute respiratory tract infection in Fuzhou, China

目的建立人冠状病毒（HCoV）-NL63、HCoV—HKU1、HCoV—OC43、HCoV-229E的实时荧光定量PCR法（FQ-PCR），调查Hc0V在福州地区急性呼吸道感染（ARTI）儿童中的流行情况。方法收集2006年10月～2007年4月、2007年10月-2。08年4月、2008年10月-2009年4月连续3个冬春季ARTI儿童鼻咽标本共538份。其中急性上呼吸道感染（AuRTI）289例，急性下呼吸道感染（ALRTI）249例。建立分别对HCoV-NL63、HCov-HKU1、HCoV—OC43、HCoV229E特异的FQ-PCR法。计数资料采用x2检验。结果建立了检测HCoV—NL63、HCoV-HKU1、HCoV—OC43、HCoV229E特异的FQ-PCR法，批内变异系数（CV）值与批间CV值均≤1．6V0。538份标本中，共检出冠状病毒阳性41份，占7．6％。其中HCoV—NL63阳性8份，占1．5％，含AURTI1例，ALRTI7例；HCoV-229E阳性5份，占0．9％，含AURTI1例，ALRTI4例；HCoV-HKU1阳性6份，占1．1％，含AURTI1例，ALRTI5例；HCoV—OC43阳性22份，占4．1％，含AuRTI13例，ALRTI9例。各型HCoV在各年份间的阳性率差异无统计学意义（P〉0．05）。HCoV—OC43阳性率显著高于HCoV—NL63、HCoV-229E和HCoV—HKU1（x2=6．721，10．979，9．387；均P〈0．01）。4种HCoV在连续3个冬春季节均有检出。结论HCoV-NL63、HCoV-HKU1、HCoV—OC43和HCoV-229E可能是冬春季福州地区ARTI患儿的病毒病原，对其进行临床检测有利于ARTI的病原学诊断与流行病学研究。

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-OC43 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%; i with AURTI, 4 withALRTI）, 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）. Conclusions It is suggested that the four eoronaviruses 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.