Evaluation of “Indigenous Absorbed ELISA Kit” for the Estimation of Seroprevalence ofMycobacterium aviumSubspeciesparatuberculosisAntibodies in Human Beings in North India

Abstract

In present pilot study aimed to estimate, presence of Mycobacterium avium subspecies paratuberculosis (MAP) antibodies in the human serum samples originating from North India using “Indigenous absorbed ELISA kit” (ELISA kit). The phase I, “ELISA kit” was optimized using protoplasmic antigen from native isolate of MAP “Indian Bison type” recovered from the biopsies of Crohn's disease patients. The phase II, sensitivity and specificity of the kit were estimated as 40.0 and 83.3%, respectively, when evaluated in 40 human serum samples (5 Crohn's disease and 22 ulcerative colitis patients and 13 healthy human subjects) with defined MAP status with respect to stool culture. Seroprevalence of MAP antibodies was higher in CD patients (80.0%) as compared to ulcerative colitis patients (4.5%) and normal human subjects (15.3%). The phase III, seroprevalence of MAP antibodies was estimated as 23.4%, on the basis of the screening of 452 human serum samples (without history) from different geographical regions of North India. Region-wise, 34.0, 33.3, 32.8, 25.0, 23.0, 17.7, and 12.5% samples were positive from the states of Punjab, Uttarakhand, New Delhi, Himachal Pradesh, Haryana, Uttar Pradesh and Jammu and Kashmir, respectively. Study reported moderately higher presence of MAP antibodies in human population, which necessitates programs to reduce the bioburden of MAP in the environment and in animal population.

First mass screening of the human population to estimate the bio-load of Mycobacterium avium subspecies paratuberculosis in North India

“…Presence of MAP in blood has been directly related with etiological role in CD (Naser et al, 2009). However, presence of MAP in blood of healthy individuals in present study has been reported earlier also (Singh et al, 2011c) has given rise to controversies and food for especially those people who contradicts the role of MAP in CD.…”

“…Previously developed "indigenous ELISA test" for other livestock species (Collin et al, 2005, Pruvot et al, 2013 has been standardized for the screening of human serum samples (Singh et al, 2011c). Using "indigenous ELISA kit" in the present investigation indicated high (34%) bioload of MAP in the human population indicating heavy exposure to MAP infection.…”

“…In a sero-survey of animal keepers suspected and not-suspected for CD showed that 12.9 and 4.2% were positive by the indigenous ELISA test (Shisodiya et al, 2009;Singh et al, 2011a) while screening of animal attendants who worked with goatherds endemic for MAP infection had higher prevalence of MAP infection as compared with person with no history of contact with animals. In another study by Singh et al (2011c), Sero-prevalence of MAP was higher in CD patients (80%), as compared to ulcerative colitis patients (4.5%) and apparently normal persons (15.3%) using "indigenous adsorbed ELISA test. The further reported sero-prevalence of MAP in another group of apparently normal human beings as 23.4%.…”

“…These findings indicated that "Indian bison type" strain may be capable of initiating disease that manifests similarly to JD in animals. "Indigenous ELISA" using PPA from "Indian bison type" biotype MAP of goat origin was superior when compared with imported commercial ELISA kits for screening of animals was also useful in screening of human samples for MAP infection (Singh et al, 2011c). Presence of MAP in patients suspected for infectious and non-infectious clinical conditions showed that besides association with Inflammatory bowel disease (Crohn"s disease), MAP may also play an important role in other health disorders and cases of colitis and other abdominal ailment in India.…”

The zoonotic potential ofMycobacterium aviumssp.paratuberculosis: a systematic review and meta-analyses of the evidence

“…Globally, human isolates were considered most likely to be of bovine origin [51][52][53][54][55], although the human isolates from two studies were also closely related to ovine type M. paratuberculosis [52,53]. In addition, two studies from India reported that a high proportion of human isolates were bison type, results so far unique to India [56,91]. Wynne et al [51] described human isolates that matched the bovine isolate from samples taken at the same time in the same region, lending evidence for potential sharing of M. paratuberculosis between species.…”