Cloning, expression, purification and serodiagnostic evaluation of fourteen Mycobacterium paratuberculosis proteins

Cho, Dong-Hee; Shin, Sung Jae; Talaat, Adel M.; Collins, Michael T.

doi:10.1016/j.pep.2006.12.022

Cited by 42 publications

(37 citation statements)

References 46 publications

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“…Conceptually, the use of specific purified proteins as the solid phase antigen in ELISA assay have suffered from low diagnostic sensitivity (Huntley et al, 2005). Recombinant antigens can also alter antigenicity due to conformational changes or lack of post-translational modifications (Cho et al, 2007;Li et al, 2007). The use of MAP CF proteins as antigen in ELISA assay increased sensitivity by 25% over commercial ELISA kits for low shedding animals (Shin et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

“…Earlier reports have identified several antigens inducing strong antibody responses but most of them have been found unsuitable for serodiagnosis because they are highly conserved within mycobacteria and hence cross-react with other mycobacterial pathogens. Recently, various MAP specific proteins/genes has been characterized, cloned, expressed and the protein was evaluated for their diagnostic potential (Cho et al, 2007). However, strain of MAP may also tune the sensitivity and specificity of test.…”

Section: Advances In Animal and Veterinary Sciencesmentioning

confidence: 99%

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Immunoreactivity to Culture Filtrate Proteins of Mycobacterium avium Subspecies paratuberculosis in Naturally Infected Goat and Sheep Sera

Gupta¹

2015

Adv. Anim. Vet. Sci.

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INTRODUCTIONM ycobacterium avium subspecies paratuberculosis (MAP), a cause of Johne's disease (Paratuberculosis) is the most wide-spread and highly prevalent due to difficulties in diagnosing pre-clinical cases. MAP has wide host range including domestic and wild ruminants, free grazing animals and also the human beings (Singh et al., 2010;Singh et al., 2014a;Singh et al., 2014b). Paratuberculosis has high economic impact on dairy industry and > USD 250 million economic losses has been reported in US alone (Ott et al., 1999). Though disease is endemic in India and economic losses in dairy farm was estimated from reproductive disorders (Rs. 23400.0/cow/year), forced removal (Rs. 41,750.0/cow/year), reduced milk yield (Rs. 5,712.0/cow/ year) and increased mortality (Rs. 11,666.0/ cow/year) (Rawat et al., 2014). Control of paratuberculosis has been hindered due to lack of efficient and accurate diagnostic tests. Lower specificity of tests can also be a problem due to the sharing of antigens or epitopes among MAP and other mycobacteria (Collins et al., 1991). Sensitivity of commercially available ELISA kits prepared using protoplasmic antigen has been reported comparatively low, 13.6-33.3% (Singh et al., 2007).Recent studies have focused on developing improved serodiagnostics using species-specific multiple protein antigens (Singh et al., 2014c). Microfluidics and Lab-on-Chip are some of the recent technologies that can predicate development of laboratory-free diagnostic devices for Research ArticleAbstract | Mycobacterium avium subspecies paratuberculosis (MAP), the cause of granulomatous chronic enteritis in ruminants ( Johne's disease) is under reported due to difficulties in diagnosing pre-clinical cases. Compromised specificity is a problem due to extensive sharing of antigens /epitopes among MAP and other mycobacterial strains. Culture filtrate (CF) proteins profile of native 'Indian Bison Type' strain of MAP was studied at different harvesting times (2-8 weeks of growth) in Middlebrook 7H9 medium supplemented with ADC, PANTA antibiotics and mycobactin J. Analysis of harvested CF proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the greater part of CF proteins had molecular masses (<70 kDa) as 14, 19, 26, 34-41, 52-55, and 70 kDa. Immunoblotting showed reactivity of CF proteins commonly recognised (28, 34-36, 38-42, 45, and 56 kDa) with all four MAP infected goat and sheep sera at 2-8 weeks of growth. Collectively, these immunoreactive MAP CF proteins could be the potential targets for developing diagnostics against Johne's disease with improved sensitivity and high specificity instead of whole cell sonicated crude protoplasmic extracts (PPA). Keywords

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Section: Discussionmentioning

confidence: 99%

Section: Advances In Animal and Veterinary Sciencesmentioning

confidence: 99%

Immunoreactivity to Culture Filtrate Proteins of Mycobacterium avium Subspecies paratuberculosis in Naturally Infected Goat and Sheep Sera

Gupta¹

2015

Adv. Anim. Vet. Sci.

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“…The use of crude antigens has been identified as a problem for the immunological diagnosis of JD in many studies. 1,[7][8][9]23,24,43 There is a need for defined specific antigens that could enable extension of the assay to identify animals likely to be immune and those with a latent infection that will eventually succumb to disease. It may enable identification of sheep that go on to develop a specific lesion type.…”

Section: Discussionmentioning

confidence: 99%

Enzyme-Linked Immunospot: An Alternative Method for the Detection of Interferon Gamma in Johne's Disease

et al. 2009

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Abstract. To date, the sensitivity of the interferon gamma (IFN-c) enzyme-linked immunosorbent assay (ELISA) to detect Johne's disease (JD) has been poor, especially in the early stages of disease. To improve the sensitivity of IFN-c detection in the early stages of infection, an alternate assay needs to be developed. The enzyme-linked immunospot (ELISPOT) assay is a highly sensitive technique for the detection of cytokines and has the potential to improve the diagnosis of JD. Of the variables examined, choice of capture antibody and the method by which the peripheral blood mononuclear cells were isolated significantly affected the ability to enumerate IFN-c-secreting cells. The ELISPOT assay was as sensitive as or better than the IFN-c ELISA at detecting ovine JD and could also detect disease at early time points postinoculation. The IFN-c ELISPOT could distinguish infected from unexposed animals; however, neither the IFN-c ELISA nor the ELISPOT assay could distinguish between sheep experimentally infected with Mycobacterium avium subspecies paratuberculosis and those exposed to the bacterium but diagnosed as uninfected at necropsy.

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“…Each of these diagnostic tests has a variable sensitivity and specificity which is crucial for both the initial screening test and confirmatory assays. All diagnostic methods are fraught with difficulties that have impeded the control and eradication of pTB (Wood et al 1989;Collins 1996;Pavlas et al 1997;Cho et al 2007;Leroy et al 2007;Martinson et al 2008;Shin et al 2008;Fern andez-Silva et al 2011;Mon et al 2012;Wadhwa et al 2012;Singh 2015). In chronic infection like pTB, local strains are key to the development of efficient diagnostic tests and vaccines; therefore, the use of commercial diagnostic kits globally has led to underreporting of the infection whenever used.…”

Section: Diagnosismentioning

confidence: 99%

“…Huge and diverse natural resources of domestic livestock species in terms of breeds and strains are found on the Earth. Such polymorphism and inherent capacity of individual animals to resist MAP infection can form the basis of breeding of animals for disease resistance as an alternative tool for building the 'herd immunity' (Romain et al 1993;Willemsen et al 2006;Cho et al 2007;Roupie et al 2008;Gioffre et al 2009;Souza et al 2011;Mon et al 2012;Roupie et al 2012;Carlos et al 2015). In addition, secretory proteins are either present in very scanty (low) amount or absent in vaccines, which are as a result unlikely to induce antibodies against the secretory proteins.…”

Section: Natural Resistance Of Animals To Ptbmentioning

confidence: 99%

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock

Chaubey

Gupta

et al. 2016

Veterinary Quarterly

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Paratuberculosis (pTB) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) in a wide variety of domestic and wild animals. Control of pTB is difficult due to the lack of sensitive, efficacious and cost-effective diagnostics and marker vaccines. Microscopy, culture, and PCR have been used for the screening of MAP infection in animals for quite a long time. Besides, giving variable sensitivity and specificity, these tests have not been considered ideal for large-scale screening of domestic livestock. Serological tests like ELISA easily detects anti-MAP antibodies. However, it cannot differentiate between the vaccinated and infected animals. Nanotechnology-based diagnostic tests are underway to improve the sensitivity and specificity. Newer generation diagnostic tests based on recombinant MAP secretory proteins would open new paradigm for the differentiation between infected and vaccinated animals and for early detection of the infection. Due to higher seroreactivity of secretory proteins vis-a-vis cellular proteins, the secretory proteins may be used as marker vaccine, which may aid in the control of pTB infection in animals. Secretory proteins can be potentially used to develop future diagnostics, surveillance and monitoring of the disease progression in animals and the marker vaccine for the control and eradication of pTB.

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Cloning, expression, purification and serodiagnostic evaluation of fourteen Mycobacterium paratuberculosis proteins

Cited by 42 publications

References 46 publications

Immunoreactivity to Culture Filtrate Proteins of Mycobacterium avium Subspecies paratuberculosis in Naturally Infected Goat and Sheep Sera

Immunoreactivity to Culture Filtrate Proteins of Mycobacterium avium Subspecies paratuberculosis in Naturally Infected Goat and Sheep Sera

Enzyme-Linked Immunospot: An Alternative Method for the Detection of Interferon Gamma in Johne's Disease

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock

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