Corrigendum to “Experimental infection model for Johne's disease using a lyophilised, pure culture, seedstock of Mycobacterium avium subspecies paratuberculosis” [Veterinary Microbiology 141 (2010) 301-311]

2018 ◽  
Vol 224 ◽  
pp. 119
Author(s):  
Douglas J. Begg ◽  
Kumudika de Silva ◽  
Lyrissa Di Fiore ◽  
Deborah L. Taylor ◽  
Katrina Bower ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Experimental Infection ◽  
Pure Culture ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Infection Model ◽  
Experimental Infection Model

scholarly journals Membrane and Cytoplasmic Proteins of Mycobacterium avium subspecies paratuberculosis that Bind to Novel Monoclonal Antibodies

2018 ◽  
Vol 6 (4) ◽  
pp. 127 ◽  
Author(s):  
John Bannantine ◽  
Judith Stabel ◽  
John Lippolis ◽  
Timothy Reinhardt
Keyword(s):  
Monoclonal Antibodies ◽  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Cell Extract ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mass Spectrometry Analysis ◽  
Lambda Phage ◽  
Expression Library ◽  
Spectrometry Analysis

Monoclonal antibodies against Mycobacterium avium subspecies paratuberculosis (Map) proteins are important tools in Johne’s disease research and diagnostics. Johne’s disease is a chronic inflammatory intestinal disease of cattle, sheep, and other ruminant animals. We have previously generated multiple sets of monoclonal antibodies (mAbs) in different studies; however, because many were generated and screened against a whole-cell extract of Map, the antigens that bind to these antibodies remained unknown. In this study, we used three different approaches to identify the corresponding Map antigens for 14 mAbs that could not be identified previously. In the first approach, a new Map-lambda phage expression library was screened to identify corresponding antigens for 11 mAbs. This approach revealed that mAbs 7C8, 9H3, 12E4, 3G5, and 11B8 all detect MAP_3404 encoding the biotin carboxylase subunit of acetyl-CoA carboxylase, while mAbs 7A6, 11F8, and 10C12 detect the GroEL2 chaperonin (MAP_3936), 6C9 detects electron transfer flavoprotein (MAP_3060c), and 14G11 detects MAP_3976, a lipoprotein anchoring transpeptidase. The epitopes to a selection of these mAbs were also defined. In a second approach, MAP_2698c bound monoclonal antibody (mAb) 14D4 as determined using protein arrays. When both of these approaches failed to identify the antigen for mAb 12C9, immunoprecipitation, mass spectrometry analysis, and codon optimization was used to identify the membrane protein, MAP_4145, as the reacting antigen. Characterized antibodies were used to quickly interrogate mycobacterial proteomic preps. We conclude by providing a complete catalog of available mAbs to Map proteins, along with their cognate antigens and epitopes, if known. These antibodies are now thoroughly characterized and more useful for research and diagnostic purposes.

Development and Standardization of Visual Loop Mediated Isothermal Amplification (LAMP) Essay for Specific Diagnosis of Johne’s Disease

2020 ◽  
Author(s):  
Manju Singh ◽  
Saurabh Gupta ◽  
Shoor Vir Singh ◽  
Gururaj Kumaresan ◽  
Deepansh Sharma ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Lamp Assay ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Isothermal Amplification ◽  
Assay Method ◽  
Loop Mediated Isothermal Amplification ◽  
Is900 Pcr

Mycobacterium avium subspecies paratuberculosis (MAP), causative agent of Johne’s disease (JD) is chronic granulomatous enteritis affecting domestic and wild ruminants. Since, MAP is not killed by pasteurization, it has been isolated from commercially pasteurized milk and milk products resulting exposure of human population to this pathogen through milk. Control and eradication of JD is considered difficult because of its insidious nature and lack of early, rapid and accurate diagnostic tests. Therefore in present study, a visual loop-mediated isothermal amplification (LAMP) assay method has been developed using a total of six primers including 2 outer (F3 and B3), 2 inner (FIP and BIP) and 2 loop (LF and LB) primers specific for MAP for the first time on ‘S 5’ strain of Mycobacterium avium subsp. paratuberculosis ‘Indian Bison type’ biotype. After laboratory standardization, final optimized reaction performed at 65°C for 45 min was achieved after titration of incubation time, temperature conditions and the reporter dye calcein. Sensitivity and specificity of the LAMP assay was optimized and compared with traditional IS900 PCR. The sensitivity of LAMP assay was found to detect 10fg (100%) of DNA and 95.7% specificity was recorded with respect to traditional IS900 PCR. Comparison showed that LAMP had 98.6% and 96.1% sensitivity and specificity of 96.1% and 92.3%, with respect to microscopy and culture exhibiting ‘Almost perfect’ strength of agreement. The study concluded that LAMP assay was a reliable and sensitive diagnostic test to detect MAP infection in feces and can also be used for the ‘mass screening’ of the milk samples with the help of less expertise.

Sign in / Sign up

Export Citation Format

Share Document