12例新型冠状病毒感染儿童病毒核酸和细胞因子检测分析

Analysis of viral nucleic acid and cytokines in 12 children with 2019-nCoV infection

目的分析新型冠状病毒(2019-nCoV)感染儿童的病毒核酸和细胞因子检测情况。方法回顾性分析2020年1—4月期间,广州市妇女儿童医疗中心收治的2019-nCoV感染确诊儿童病例的临床和实验室资料。隔日采集咽拭子和肛拭子,荧光定量PCR法检测咽拭子和肛拭子2019-nCoV核酸。流式细胞术检测病程早期(咽拭子和肛拭子病毒核酸检测均为阳性)、中期(咽拭子病毒核酸阴性而肛拭子核酸仍阳性)、恢复期(咽拭子和肛拭子病毒核酸检测均阴性)血清细胞因子IFN-γ/IL-1β/IL-2/IL-4/IL-5/IL-6/IL-8/IL-10/IL-12p70/IL-17A/IL-17F/IL-22/TNF-α/TNF-β水平。结果共12例患儿纳入本研究。男∶女=5∶1;平均年龄(7.0±4.3)岁;临床分型无症状感染2例,轻型5例,普通型5例,无重症及危重症患者。9例我院初诊患儿咽拭子和肛拭子病毒核酸最初呈平行阳性状态,肛拭子排毒中位数时间长于咽拭子排毒时间[32(4.5,45.0)d vs 3(2,9)d,Z=11.0,P=0.010];肛拭子较咽拭子排毒时间差值中位数长25.5(1.5,42.8)d。病程不同时期(早期、中期、恢复期)血清细胞因子IL-17A、IL-4和IL-5总体水平比较差异有统计学意义(Z或F,P值分别为8.33,0.016;5.36,0.010;6.56,0.004),但主要在病程中期明显升高。而IL-17F、IL-2、IL-22在病程的早、中、恢复期均有升高,但病程各组间比较差异无明显统计学意义(P>0.05)。结论肠道排毒时间较长,尽管尚未确定感染性,但对防控提出了更高要求。Th2细胞和Th17细胞分泌的细胞因子参与了2019-nCoV感染后的非重症感染儿童的免疫应答过程,监测儿童患者的病毒排出情况和细胞因子变化,对于判断患者的病情有较好的参考价值。

Objective To analyze the viral nucleic acid and cytokines in 12 children with 2019-nCoV infection.Methods Clinical and laboratory data of the children diagnosed with 2019-nCoV infection in Guangzhou Women and Children′s Medical Center from January to April 2020 were retrospectively analyzed.Throat and anal swabs were collected on alternate days for the detection of 2019-nCoV nucleic acid by fluorescence quantitative PCR.Flow cytometry was used to detect serum cytokines including IFN-γ,IL-1β,IL-2,IL-4,IL-5,IL-6,IL-8,IL-10,IL-12p70,IL-17A,IL-17F,IL-22,TNF-αand TNF-βduring the early(both throat and anal swab tests were positive),the intermediate(throat swab test was negative,while anal swab test remained positive),and the convalescence(both throat and anal swab tests were negative)stages of infection.Results A total of 12 children were enrolled in this study.The male-to-female ratio was 5∶1.The average age was(7.0±4.3)years.There were two asymptomatic,five mild and five common cases.No severe or critical cases were involved.Initially,throat and anal swab nucleic acid tests were simultaneously positive in nine children newly diagnosed in our hospital and the median time of viral shedding in throat swab was longer than that in throat swab[32(4.5,45.0)d vs 3(2,9)d,Z=11.0,P=0.010].The median difference of viral shedding time between anal swab and pharyngeal swab was 25.5(1.5,42.8)d.The overall levels of serum cytokines IL-17A,IL-4 and IL-5 in different stages of the disease(early,intermediate and convalescence stage)were statistically different(Z or F,P values were 8.33,0.016;5.36,0.010 and 6.56,0.004,respectively),and a significant increase was observed in the intermediate stage of infection.IL-17F,IL-2 and IL-22 were all increased during the infection,but there was no significant statistical difference among the three stages(P>0.05).Conclusions It was noted that intestinal viral shedding needed a longer time.Although the infectivity has not been determined,higher requirements have been put forward for disease prevention and control.Cytokines secreted by Th2 and Th17 cells were involved in the immune response in children with non-severe 2019-nCoV infection.Monitoring viral shedding and cytokine changes in pediatric patients would be conducive to disease assessment.