Real-time PCR versus traditional and Nano-based ELISA in early detection of murine trichinellosis

Abstract

Abstract Trichinellosis is a serious foodborne zoonosis. It poses a serious risk to public health worldwide. Early serological diagnosis of trichinellosis is influenced by an immunological ‘silent’ phase following infection. This highlights the necessity for developing sensitive diagnostic approaches to be employed when antibodies cannot be detected. In this work, the validity of traditional ELISA, Nano-ELISA and real time polymerase chain reaction (PCR) were evaluated in early diagnosis of Trichinella spiralis. Swiss albino mice were orally infected with 100 and 300 muscle larvae/mouse. Mice were sacrificed 4, 6, 8, 10, 15, and 28 days post-infection (dpi). Blood samples were tested for circulating antigen by traditional ELISA and Nano-ELISA using anti-rabbit polyclonal IgG conjugated with AgNPs and for Rep gene by SYBR green real-time PCR. Rep gene detection by SYBR green real-time PCR could detect T. spiralis with 100% sensitivity in the mild infection group at 8 dpi, while in the severe infection group it reached 100% sensitivity at 4 dpi. Nano-ELISA could detect T. spiralis circulating antigen from 4 dpi in both mild and severe infection and reached 100% sensitivity at 8 dpi and 6 dpi in mild and severe infection, respectively. However, traditional ELISA could detect T. spiralis circulating antigen from 6 dpi and reached maximum sensitivity at 15 dpi in the mild infection group, while in the severe infection group detection began at 4 dpi and reached 100% sensitivity at 8 dpi. Nano-ELISA and real time PCR, using Rep gene, are useful tools for the detection of early T. spiralis infection even in its mild infection state.