Prevalence of Mouse Norovirus in Experimental Mice in Beijing

doi:10.12300/j.issn.1674-5817.2022.126

Abstract

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

CLC Number:

Q95-33

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.

Figures/Tables 5

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

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.

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

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

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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