北京地区实验小鼠诺如病毒流行情况

doi:10.12300/j.issn.1674-5817.2022.126

实验动物与比较医学 ›› 2023, Vol. 43 ›› Issue (2): 205-212.DOI: 10.12300/j.issn.1674-5817.2022.126

北京地区实验小鼠诺如病毒流行情况

刘芳妮¹^,²(), 吕军苹²^,³, 柯跃华², 王长军¹^,², 郭金鹏²()()

^1.中国医科大学公共卫生学院, 辽宁 110122
^2.解放军疾病预防控制中心应急处置大队科室, 北京 100071
^3.中央民族大学药学院, 北京 100081

收稿日期:2022-08-16 修回日期:2022-12-18 出版日期:2023-04-25 发布日期:2023-05-16
通讯作者: 郭金鹏（1977—），男，博士，副研究员，研究方向：军事预防医学。E-mail： pengpla@126.com。ORCID：0000-0001-5966-4109
作者简介:刘芳妮（1994—），女，硕士研究生，研究方向：诺如病毒感染机制研究。E-mail： fangniliu@163.com
基金资助:
军队后勤生物安全研究专项“生物样本多途径核酸及血清学调查技术研究”(2019T03901)

Prevalence of Mouse Norovirus in Experimental Mice in Beijing

Fangni LIU¹^,²(), Junping LU²^,³, Yuehua KE², Changjun WANG¹^,², Jinpeng GUO²()()

^1.School of Public Health, China Medical University, Liaoning 110122, China
^2.Emergency response brigade department, PLA Center for Disease Control and Prevention, Beijing 100071, China
^3.college of pharmacy, Minzu University of China, Beijing 100081, China

Received:2022-08-16 Revised:2022-12-18 Published:2023-04-25 Online:2023-05-16
Contact: GUO Jinpeng (ORCID: 0000-0001-5966-4109), E-mail: pengpla@126.com

摘要/Abstract

摘要：

目的调查北京地区19家生物公司自然条件下养殖的实验小鼠感染小鼠诺如病毒（mouse norovirus，MNV）的情况。方法从19家公司生产的小鼠中随机抽取，每品种品系、每批次14只小鼠合为一笼，共计1 396笼19 544只小鼠。对每笼取粪便样本，采用TaqMan探针荧光定量PCR法，用MNV-1引物检测各品系小鼠感染MNV的情况，根据循环阈（cycle threshold，Ct）值判断小鼠是否被感染；采用卡方检验对5种小鼠粪便样本阳性率进行统计学差异性分析；将5种小鼠阳性样本的Ct值进行统计描述；采用非参数检验法对5种小鼠Ct值进行差异性分析。结果共检测粪便样本1 396份，BALB/c、C57BL/6、ICR、KM、BALB/c-nude小鼠的粪便MNV检出阳性率分别为17.65%、39.33%、10.57%、18.32%和27.4%。 C57BL/6小鼠的粪便MNV检出阳性率大于BALB/c、ICR、KM小鼠（P＜0.05），且BALB/c -nude小鼠的粪便MNV检出阳性率大于ICR、BALB/c小鼠（P＜0.05）；BALB/c-nude小鼠和C57 BL/6小鼠的病毒载量总体上大于KM小鼠（P＜0.001，P＜0.05）。结论 MNV-1引物可应用于小鼠MNV感染情况的检测；北京市5种实验小鼠MNV阳性率在10%～40%，其中C57BL/6小鼠和BALB/c -nude小鼠的MNV阳性检出率较高。

关键词: 小鼠诺如病毒, TaqMan探针, 荧光定量PCR, 自然感染

Abstract:

Objective To investigate the infection of mouse norovirus (MNV) in experimental mice raised under natural conditions from 19 biological companies in Beijing. Methods The mice used in this study were randomly selected from mice produced by 19 companies, and 14 mice of each strain and batch were combined into one cage, totaling 1 396 cages of 19 544 mice. The fecal samples from BALB/c, C57BL/6, ICR, KM, and BALB/c-nude mice were collected. TaqMan probe fluorescence quantitative PCR method was used to detect MNV infection of mice with MNV-1 primer, and whether the mice were infected with MNV was determined according to cycle threshold (Ct value). The chi-square test was used to analyze the difference of positive rate among the fecal samples from the five types of mice. The Ct values of the positive samples were statistically described; the non-parametric test was used to analyze the differences in Ct values among the five types of mice. Results A total of 1 396 fecal samples were collected. The positive rates of fecal MNV detection in BALB/c, C57BL/6, ICR, KM, and BALB/c-nude mice were 17.65%, 39.33%, 10.57%, 18.32% and 27.4%, respectively. According to the chi-square test results, the positive rate of fecal in C57BL/6 mice was higher than that in BALB/c, ICR, and KM mice (all P<0.05), and the positive rate of BALB/c-nude mice was higher than that in ICR and BALB/c mice (P<0.001, P＜0.05) . The viral load of BALB/c-nude or C57BL/6 mice was generally greater than that of KM mice (P<0.05). Conclusion MNV-1 primers can be applied to the detection of MNV infection in mice. The positive rate of MNV in five types of experimental mice in Beijing ranges from 10% to 40%, among which C57BL/6 mice and BALB/c-nude mice have higher positive rates of MNV than the others.

Key words: Mouse norovirus, TaqMan probe, Fluorescence quantitative PCR, Natural infection

中图分类号:

Q95-33

刘芳妮, 吕军苹, 柯跃华, 王长军, 郭金鹏. 北京地区实验小鼠诺如病毒流行情况[J]. 实验动物与比较医学, 2023, 43(2): 205-212.

Fangni LIU, Junping LU, Yuehua KE, Changjun WANG, Jinpeng GUO. Prevalence of Mouse Norovirus in Experimental Mice in Beijing[J]. Laboratory Animal and Comparative Medicine, 2023, 43(2): 205-212.

图/表 5

图1 MNV-1引物的实时荧光PCR标准曲线图注：Ct是循环阈值。

Figure 1 The Standard curve of real-time PCR for MNV-1 primerNote： Ct， cycle threshold.

图2 TaqMan荧光定量PCR检测MNV特异性的结果注：2、6号为小鼠诺如病毒；其余病毒依次为1号ev71、3号鼠肝炎病毒、4号冠状病毒229e、5号脊髓灰质炎病毒、7号柯萨奇病毒、8号埃可病毒、9号腺病毒、10号甲流病毒、11号乙流病毒、12号甲肝病毒；N为阴性对照。基线值为FAM：1 808 874。

Figure 2 Results of specificity test of the TaqMan real-time RT-PCR assay for murine noroviruses （MNV）Note： No.2 and No.6 are murine noroviruses; The other viruses are as follows: No.1 is ev71, No.3 is murine hepatitis virus, No.4 is Coronavirus 229e, No.5 is poliovirus, No.7 is coxsackie virus, No.8 is echovirus, No.9 is influenza A virus, No.11 is influenza B virus, No.12 is hepatitis A virus. N: negative control. The baseline value was FAM:1 808 874.

表1 北京市小鼠粪便样本的MNV病毒检出情况

Table 1 Detection of MNV in mouse fecal samples in Beijing

小鼠品种品系 Stock or strain of mice	样本数 Samples	阳性数量 Positive samples	阳性率 Positive rate/%
BALB/c	425	75	17.65^△
C57BL/6	267	105	39.33
ICR	265	28	10.57^*△
KM	202	37	18.32^*
BALB/c-nude	237	65	27.40^*
总样本数量 The total number of sample	1 396	310	22.21

图3 小鼠Ct值频数分布直方图注：“［”表示包含端点值，“）”表示不包含端点值。

Figure 3 Histogram of frequency distribution of Ct values of miceNote：“［“ represents inclusion of endpoint value; ”) “ represents exclusion of endpoint value.

图4 5个品种品系小鼠阳性样本Ct值箱式图注：经Kruskal-Wallis检验的多重比较以及Bonferrni校正，***P＜0.001，*P＜0.05

Figure 4 The Box plot of Ct values for positive samples of five stocks or strains of miceNote：After K-W test for multiple comparisons and Bonferrni correction， ***P <0.001， *P <0.05.

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北京地区实验小鼠诺如病毒流行情况

Prevalence of Mouse Norovirus in Experimental Mice in Beijing

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