Development of a loop-mediated isothermal amplification (LAMP) assay targeting the mitochondrial cytochrome b gene for the rapid detection of alveolar echinococcosis in hepatic nodules of horses.

The study presents the development of a rapid detection tool called the Loop-mediated Isothermal Amplification (LAMP) assay, which targets the mitochondrial cytochrome b gene to diagnose alveolar echinococcosis in horses’ hepatic nodules. The technique was proven equally effective as the conventional Polymerase Chain Reaction (PCR) testing.

Objectives of the Study

• The researchers aim to improve early detection methods for alveolar echinococcosis in horses because of their importance to public health. This disease can be contracted by humans and is caused by a parasitic infection of Echinococcus multilocularis.
• The study seeks to develop a highly specific and sensitive diagnostic method that can be used during meat inspections, as alveolar echinococcosis has been reported in slaughtered horses in Japan and Poland.

Methodology

• The study involved developing a LAMP assay targeted at the mitochondrial cytochrome b (cob) gene of the E. multilocularis parasite.
• Forty-one hepatic nodules from horses were examined histopathologically and submitted to a cob-targeted PCR test and the newly developed LAMP assay.
• The researchers tested the LAMP assay under optimal conditions, which were determined as 64℃ (degree Celsius) for 30 minutes.

Results and Findings

• The results from the LAMP assay were found to be consistent with those obtained from the cob-targeted PCR.
• The detection limit of the cob gene using the LAMP assay was found to be 135 copies/μL.
• The findings suggest that the LAMP assay’s ability is equivalent to conventional PCR testing. Therefore, it presents an alternate method for routine genetic diagnosis of alveolar echinococcosis in horses.

Implications and Conclusions

• The successful development and validation of the LAMP assay for rapid detection of alveolar echinococcosis bring an alternative to the conventional PCR. The LAMP assay can provide a rapid, cost-effective, and efficient diagnosis during meat inspections, reducing the risk of zoonotic transmission to humans.
• Considering the reported cases of alveolar echinococcosis from slaughtered horses in Japan and Poland, the application of the LAMP assay technique could be significant in ensuring public health safety in these specific regions and potentially worldwide.