2009至2010年南京儿童医院5岁以下儿童病毒性腹泻分子流行病学特点

Molecular epidemiological study on viral diarrhea among pediatric patients under five years old in Nanjing,2009-2010

目的对南京儿童医院(我院)5岁以下儿童轮状病毒(HRV)、人杯状病毒(HuCV)、星状病毒(AstV)和肠道腺病毒(AdV)感染的分子流行病学进行研究,为病毒性腹泻的防治提供基础数据和理论依据。方法收集2009年7月至2010年6月于我院消化科门诊就诊的5岁以下腹泻患儿的粪便标本。采用ELISA法检测A组HRV,阳性标本采用RT-PCR法进行毒株分型鉴定;HuCV、AstV和AdV采用RT-PCR或PCR法进行检测。结果研究期间共收集病毒性腹泻患儿粪便标本300份,其中男188例,女112例。年龄1～59月龄,平均(10.5±9.2)月龄。4种病毒检测阳性率依次为HRV(37.7%,113/300)、HuCV(23.7%,71/300)、AstV(4.0%,12/300)和AdV(2.7%,8/300)。①HRVG血清型以G3型(38.9%)最常见,其次为G2型(8.8%);P基因型以P[8](33.6%)为主。G血清型和P基因型组合以G3P[8](15.9%)为主。113份HRV阳性标本中,7～12月龄儿童占43.4%(49/113),高发季节为2009年10月至2010年1月。②HuCV感染中诺如病毒检出67份,扎如病毒4份。HuCV感染于2009年8月出现一个小高峰,发病年龄高峰为7～12月龄(38/71),24月龄以下患儿占95.8%(68/71)。③10/12例AstV感染发生于2009年10月至2010年1月,月龄分布为2～16月龄。④8例AdV阳性标本PCR产物经克隆测序,AD2、3、5、7、12和41型各1例,AD31型2例。AdV感染高发于2010年1至6月,发病高峰为7～12月龄(4/8)。⑤60.0%(180/300)至少检出4种病毒中的一种。混合感染23份,其中12份(52.2%)为HRV+HuCV,5份(21.7%)为HRV+AstV,2份为HuCV+AstV,HRV+AdeV、HuCV+AdeV、AdeV+AstV、HRV+AstV+HuCV各1份。结论 HRV是引起婴幼儿病毒性腹泻最主要的病毒病原,以G3P[8]为主要优势株。HuCV、AstV和AdV也是重要的病原。

Objective A molecular epidemiological study on rotavirus （HRV）, human calicivirus （HuCV）, astrovirus （AstV） and adenovirus （AdV） was conducted in stool specimen from patients with diarrhea under 5 years of age in Nanjing Children Hospital of Nanjing Medical University from July 2009 to June 2010, to provide the basic data and theory evidence for prevention and treatment of viral diarrhea. Methods In Nanjing Children＇s Hospital of Nanjing Medical University from July 2009 to June 2010, 300 stool specimens were collected from outpatients under 5 years of age diagnosed as diarrhea. HRV was detected by ELISA, strain characterization was carried out by nested PCR. HuCV （including norovirus and sapovirus） , AstV and AdV were detected by RT-muhiplex PCR or PCR. Results In the 300 children whose samples were collected, 188 patients were boys, 112 were girls. The age of children ranged from 1 to 59 months （mean ＋ SD age： 10. 5 ＋9. 2 months）. At least one of the four viral agents was found in 60. 0% （180/300） of the specimens. HRV, HuCV, AstV and AdV were identified in 37.7% （ 113/300）, 23.7% （71/300） , 4. 0% （12/300）, and 2. 7% （8/300） in 300 specimens respectively. （~）HRV G3 （38. 9% ） was the mostprevailing serotype, followed by G2 （8.9%）. Based on P typing, P[81 （33.6%） was the most common genotype. The most common G-P combination was G3P[ 8 ] （15.9%）. The incidence of HRV infections peaked from October 2009 to January 2010, HRV was mostly found in children aged from 7 to 12 months. （~）Of 71 HuCV positive samples, norovirus was detected in 67 cases and sapovirus was detected in 4 cases. HuCV infection peaked in August 2009, and was mostly found in children aged from 7 to 12 months. 95.8% （68/71） children aged under 24 months. @Ten of 12 AstV infections occurred from October 2009 to January 2010, and AstV infected children were aged from 2 to 16 months. （~）Of 8 AdV positive samples, there were seven serotypes： Ad31 （2）, Ad2（ 1 ）, Ad3（ 1 ）, Ad5（ 1 ）, Ad7（ 1 ）, Adl2（ 1 ） and Ad41 （ 1 ）. AdV infection peaked from January to June in 2010 and was mostly found in children aged from 7 to 12 months. @Twenty-three of 300 samples were found positive to more than one viral agent, in which 12 samples contained both HRV and HuCV , and 5 samples contained both HRV and AstV, 1 sample contained both HRV and AdV, 2 samples contained both HuCV and AstV, ! sample contained both HuCV and AdV, 1 sample contained both AstV and AdV, and 1 sample contained HRV, AstV and HuCV. Conclusions The results indicated clearly the impact of viral agents in causing diarrhea in children and the importance of long-term systematic surveillance. HRV was also the most important pathogen, but HuCV, AstV and AdV should be given more attention.