Serological detection of Mycobacterium Tuberculosis complex infection in multiple hosts by One Universal ELISA.

Liang Sun,Qiaoying Zeng,Yingyu Chen,Ping Yi,Li Yang,Yu Yan,Kailun Zhang,Aizhen Guo,Simon Clegg

doi:10.1371/journal.pone.0257920

Abstract

Tuberculosis (TB), a contagious disease mainly caused by Mycobacterium tuberculosis (M. tb), Mycobacterium bovis (M. bovis), and Mycobacterium caprae (M. caprae), poses a major global threat to the health of humans and many species of animals. Developing an ante-mortem detection technique for different species would be of significance in improving the surveillance employing a One Health strategy. To achieve this goal, a universal indirect ELISA was established for serologically detecting Mycobacterium tuberculosis complex infection for multiple live hosts by using a fusion protein of MPB70, MPB83, ESAT6, and CFP10 common in M. tb, M. bovis, and M. caprae as the coating antigen (MMEC) and HRP-labeled fusion protein A and G as a secondary antibody. After testing the known positive and negative sera, the receiver operating characteristic curves were constructed to decide the cut-off values. Then, the diagnostic sensitivity and specificity of MMEC/AG-iELISA were determined as 100.00% (95% CI: 96.90%, 100.00%) and 100.00% (95% CI: 98.44%, 100.00%) for M. bovis infection of cattle, 100.00% (95% CI: 95.00%, 100.00%) and 100.0% (95% CI: 96.80%, 100.00%) for M. bovis infection of sheep, 90.74% (95% CI: 80.09%, 95.98%) and 98.63% (95% CI: 95.14%, 99.76%) for M. bovis infection of cervids, 100.00% (95% CI: 15.81%, 100.00%) and 98.81% (95% CI: 93.54%, 99.97%) for M. bovis infection of monkeys, 100.00% (95% CI: 86.82%, 100.00%) and 94.85% (95% CI: 91.22%, 97.03%) for M. tb infection of humans. Furthermore, this MMEC/AG-iELISA likely detects M. caprae infection in roe deer. Thus this method has a promising application in serological TB surveillance for multiple animal species thereby providing evidence for taking further action in TB control.

Highlights

Tuberculosis (TB) is known as an important zoonotic infectious disease caused by various Mycobacterium tuberculosis complex (MTBC) members
The main members of MTBC include Mycobacterium tuberculosis (M. tb), Mycobacterium bovis (M. bovis), and Mycobacterium caprae (M. caprae), which share a wide range of hosts but maintain different host preferences [1]
All the pre-diluted sera were tested with MMEC/AG-iELISA to further determine the parameters of this assay

Summary

Introduction

Tuberculosis (TB) is known as an important zoonotic infectious disease caused by various Mycobacterium tuberculosis complex (MTBC) members. Sheep TB cases are mainly caused by M. caprae and M. bovis transmitted from cattle under the condition of cohabitation or sharing grazing land. In Spain, sheep cohabiting with infected cattle or goats were reported to have a prevalence of 50.44% caused by the infection with M. bovis or M. caprae [13]. Wild animals such as deer and monkeys are susceptible to these three MTBC members [14,15]. OIE added the infection of bovids, cervids, goats, and New World camelids by M. bovis, M. tb, and M. caprae into the disease list of the Terrestrial Animal Health Code since 2019. To quickly investigate the epidemiological status of TB in many different species of live animals, it is ideal to have one common universal test for all

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