Multiplex Immunochromatographic Assay for Serologic Diagnosis of Major Infectious Diseases in Laboratory Mice

Abstract

Serologic monitoring of infectious diseases is important for microbial control in colonies of laboratory mice. Rapid and simple tests that do not require killing animals are valuable for this purpose. In this study, we developed a multiplex immunochromatographic assay (ICA) for detection of antibodies to mouse hepatitis virus (MHV), Sendai virus (also known as hemagglutinating virus of Japan [HVJ]), and Clostridium piliforme (The pathogen that causes Tyzzer disease), which are major infectious diseases in mice. For this assay, an ICA strip was put into a microtube containing 150 μL PBS and either 0.75 μL mouse serum or 1.5 μL whole blood. Binding antibodies were visualized by using protein A-conjugated colloidal gold. Under these conditions, multiplex ICA simultaneously and specifically detected antibodies to multiple antigens. To evaluate the sensitivity and specificity of multiplex ICA, positive serum samples for each infectious disease were used. Sensitivities of the multiplex ICA test for MHV, HVJ, and C. piliforme were 100%, 100%, and 90%, respectively. No nonspecific reaction was observed in any of the 30 positive sera. In addition, 10 samples of uninfected sera did not show any bands except for the control line. These observations indicate high specificity of the multiplex ICA test. Moreover, the multiplex ICA could be applied to diluted blood. These results indicate that the multiplex ICA is appropriate for rapid, simple, and safe serologic testing of laboratory mice.

Establishment and application of a loop‐mediated isothermal amplification−lateral flow dipstick (LAMP–LFD) method for detecting Clostridium piliforme

“…Niepceron and Licois ( 2010 ) established a nested PCR method with high sensitivity for the detection of C. piliforme ; Tosa et al. ( 2019 ) established a multiplex immunochromatographic method for rapid and simple detection of multiple pathogens, including C. piliforme . Rapid, instant and accurate nucleic acid detection is necessary in certain industries; however, some centres are not equipped with complex instruments, such as a PCR system (Wang et al., 2023 ; Yan et al., 2023 ).…”

Simultaneous serodetection of major rat infectious pathogens by a multiplex immunochromatographic assay

“…A multiplex microbead immunoassay and PCR-based detection of antigen in feces have been reported as methods for diagnosing infectious disease without euthanizing animals [1][2][3][4][5][6]. We previously reported that the use of a multiplex immunochromatographic assay (ICA) enabled simultaneous and specific detection of antibodies to mouse hepatitis virus (MHV), Sendai virus (also known hemagglutinating virus of Japan, HVJ), and Clostridium piliforme (the pathogen that causes Tyzzer disease) with small amounts of blood that can be collected without euthanizing mice [7]. Additionally, multiplex ICA is useful for in-house monitoring due to its ability to simply and rapidly detect antibodies against multiple antigens without the need for an expensive, dedicated analyzer.…”