广东省科技计划项目（2014B070706006）。
目的 了解我省小鼠腺病毒（Mad）自然感染情况及探究人工感染Mad小鼠体内各脏器组织中病毒分布规律及血清抗体变化。方法 采用ELISA方法对2007年和2015年饲养于普通环境小鼠血清及2013年~2015年广东省12家监督单位和32家委托单位SPF级小鼠血清进行病毒抗体检测。利用腹腔注射方法感染36只3周龄BALB/c小鼠，0.2 mL/只，浓度为4.5×106 copies/μL，每天观察动物临床表现，并于攻毒前第0天和攻毒后第3、7、10、15、18、21、30、37、44、51、60天剖检小鼠（各3只），取组织标本（心、肝、脾、肺、肾、脑、胃、盲肠内容物）及血清，应用实时荧光定量PCR（QPCR）方法检测组织病毒核酸，ELISA方法检测血清抗体水平。结果 普通饲养小鼠抗体阳性率为24.44%~84.15%，其中以Mad-2型K87株血清型为主。SPF级小鼠Mad血清抗体阳性率均为0%。所有攻毒小鼠均呈隐性感染，无临床表现。攻毒后第3天及7天小鼠各组织病毒核酸检测阳性率均为100%（3/3），其中以脾脏100%阳性率维持时间最长（60 d）。除肝脏外，小鼠各组织病毒含量均在攻毒后7 d达到高峰，其中以脾脏病毒含量最高（5.5×105 copies/μL），其次是心脏（3.4×105 copies/μL）、盲肠内容物（2.6×105 copies/μL）和胃（2.6×105 copies/μL），脑为0.8×105 copies/μL。攻毒后15 d可测出血清抗体，在37 d达到峰值，此后至60 d一直维持高水平。结论 SPF级小鼠Mad感染率低，普通环境饲养小鼠Mad感染率高。人工感染小鼠脾脏病毒含量最高，阳性率维持时间最长，说明病毒可以在脾脏内长期复制，感染后第7天为组织病毒核酸最佳检测时间点。血清抗体在感染后15~60 d内均可以作为监测指标。
Objective To investigate the natural infection rate of Mouse Adenovirus(Mad)in Guangdong province and to estimate the tissue distribution and serology changes of Mad in infected mice. Methods Serums of mice from general environment of Guangzhou in 2007 and 2015, and the SPF mice from12 supervision and 32 entrusted units of Guangdong province among 2013~2015 were detected by ELISA.Thirty-six 3 weeks old BALB/c mice were each intraperitoneally inoculated with 0.2 mL of Mad in 4.5×106 copies/μL concentration. The tissue samples, including heart, liver, spleen, lung, kidney, brain, stomach, cecum contents and serum were taken at 0 d before inoculation, and 3, 7, 10, 15, 18, 21, 30, 37, 44, 51, 60 d after inoculation (3 animals each time).Real-time fluorescent quantitative PCR (Q-PCR) method was used to detect the virus nucleic acid of tissue. In addition, the antibody against Mad was detected by ELISA.Results Antibody positive rate was 24.44%~84.15% and 0% among the general environment mice and the SPF mice, respectively, which is given priority to with Mad-2 K87 serotype strains. There were no clinical manifestations among all infected mice.The virus nucleic acid positive rate was 100% (3/3) in all tissue samples in 3 d and 7 d, which maintained a longest time to 60 d in spleen. Besides the liver, virus nucleic acid content were highest at 7 d in each tissue of infected mice, with spleen highest (5.5×105 copies/μL), heart was second at 3.4×105 copies/μL, cecum contents and stomach ranked third (2.6×105 copies/μL), followed by brain (0.8×105 copies/μL). The antibody could be successfully detected at 15 d after inoculated, hit the peak at 37 d and maintained a high level until 60 d.Conclusions Mad infection rate is low in SPF mice, high in general environment mice.Mad nucleic acid content is the highest and positive rate maintain the longest in spleen,suggesting that Mad mainly replicate in spleen. And 7 d is the best pot for nucleic acid testing.Serum neutralizing antibody can be used as a monitoring index within 15~60 d after infection.
罗银珠,张钰,潘金春,王静,袁文,何丽芳,吴瑞可,黄碧洪,郭鹏举,黄韧.广东小鼠腺病毒血清学调查及病毒和抗体在人工感染小鼠体内消长规律研究[J].中国比较医学杂志,2017,27(1):43~48.
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