scholarly journals A Novel Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycobacterium avium subsp. paratuberculosis Infections (Johne's Disease) in Cattle

2006 ◽  
Vol 13 (5) ◽  
pp. 535-540 ◽  
Author(s):  
C. A. Speer ◽  
M. Cathy Scott ◽  
John P. Bannantine ◽  
W. Ray Waters ◽  
Yasuyuki Mori ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Mycobacterium Avium ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Formaldehyde Concentration ◽  
Serum Samples ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Diagnostic Sensitivity And Specificity

ABSTRACT Enzyme-linked immunosorbent assays (ELISAs) for the diagnosis of Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis, were developed using whole bacilli treated with formaldehyde (called WELISA) or surface antigens obtained by treatment of M. avium subsp. paratuberculosis bacilli with formaldehyde and then brief sonication (called SELISA). ELISA plates were coated with either whole bacilli or sonicated antigens and tested for reactivity against serum obtained from JD-positive and JD-negative cattle or from calves experimentally inoculated with M. avium subsp. paratuberculosis, Mycobacterium avium subsp. avium, or Mycobacterium bovis. Because the initial results obtained from the WELISA and SELISA were similar, most of the subsequent experiments reported herein were performed using the SELISA method. To optimize the SELISA test, various concentrations (3.7 to 37%) of formaldehyde and intervals of sonication (2 to 300 s) were tested. With an increase in formaldehyde concentration and a decreased interval of sonication, there was a concomitant decrease in nonspecific binding by the SELISA. SELISAs prepared by treating M. avium subsp. paratuberculosis with 37% formaldehyde and then a 2-s burst of sonication produced the greatest difference (7×) between M. avium subsp. paratuberculosis-negative and M. avium subsp. paratuberculosis-positive serum samples. The diagnostic sensitivity and specificity for JD by the SELISA were greater than 95%. The SELISA showed subspecies-specific detection of M. avium subsp. paratuberculosis infections in calves experimentally inoculated with M. avium subsp. paratuberculosis or other mycobacteria. Based on diagnostic sensitivity and specificity, the SELISA appears superior to the commercial ELISAs routinely used for the diagnosis of JD.

scholarly journals The collection of lymphatic fluid from the bovine udder and its use for the detection of Mycobacterium avium subsp. paratuberculosis in the cow

2011 ◽  
Vol 24 (1) ◽  
pp. 23-31 ◽  
Author(s):  
Johannes L. Khol ◽  
Pablo J. Pinedo ◽  
Claus D. Buergelt ◽  
Laura M. Neumann ◽  
Walter Baumgartner ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Fluid Collection ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lymph Fluid ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Lymphatic Fluid ◽  
Elisa Result

The objective of the current study was to evaluate the feasibility of lymph collection from the bovine udder and to investigate if the lymphatic fluid might be of diagnostic value in cows infected with Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of paratuberculosis. Lymph fluid collection was attempted from 58 cows, and the reactions of the cows as well as the level of difficulty of the procedure were recorded in 56 animals. Lymph samples (51 in total) were tested for the presence of MAP by nested polymerase chain reaction. Collection of the lymphatic fluid caused no or mild signs of discomfort in 94.6% of the cows; in 51.8% of cows, lymphatic fluid was attained on the first attempt, while sample collection was unsuccessful in 12.1%. Mycobacterium avium subsp. paratuberculosis was detected in 43.1% of all lymph samples. The bacterium was present in 66.7% of cows with clinical Johne’s disease, in 42.8% of asymptomatic cows with a positive or suspicious enzyme-linked immunosorbent assay (ELISA) result in blood, and in 38.7% of cows with a negative ELISA result in blood. The present study shows that the procedure was well tolerated by most cows and can easily be performed on farm. The current report of the isolation of MAP from lymph fluid suggests that the present approach could be used for the early detection of Johne’s disease in cattle.

scholarly journals Low rate of detectable in utero transmission of Mycobacterium avium subspecies paratuberculosis in a dairy herd with a low prevalence of Johne’s disease

2011 ◽  
Vol 24 (1) ◽  
pp. 153-155 ◽  
Author(s):  
John M. Adaska ◽  
Robert H. Whitlock
Keyword(s):  
Mycobacterium Avium ◽  
Transmission Rate ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Dairy Herd ◽  
In Utero ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Causal Organism ◽  
Mycobacterium Avium Subsp Paratuberculosis

In an effort to correlate the likelihood of in utero transmission of Mycobacterium avium subsp. paratuberculosis (MAP), the causal organism of Johne’s disease, with the test status of the dam, tissues from neonatal calves borne to known test status cows were cultured for the presence of MAP. Tissues from a single calf borne to a test-positive cow shedding large numbers of organisms in the feces were positive for MAP. The detected overall transmission rate was approximately 2% (1/49), and the detected transmission rate in cows that were fecal culture positive and serum enzyme-linked immunosorbent assay suspect or positive was approximately 4.3% (1/23).

scholarly journals Improved Immunodiagnosis of Cystic Hydatid Disease by Using a Synthetic Peptide with Higher Diagnostic Value Than That of Its Parent Protein, Echinococcus granulosus Antigen B

2000 ◽  
Vol 38 (11) ◽  
pp. 3979-3983 ◽  
Author(s):  
Gualberto González-Sapienza ◽  
Carmen Lorenzo ◽  
Alberto Nieto
Keyword(s):  
Hydatid Disease ◽  
Synthetic Peptides ◽  
Diagnostic Value ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Cystic Hydatid Disease ◽  
Antigen B ◽  
Parent Protein ◽  
Diagnostic Sensitivity And Specificity

The assays are used for the diagnosis of hydatid disease are still imperfect. The reported diagnostic sensitivity and specificity vary greatly depending on the panel of sera used, the laboratory conducting the assay, and, more critically, the antigen used. To contribute to its standardization, we have recently ranked the diagnostic performances of the major parasite antigens and the available synthetic peptides using a large collection of serum samples. That work showed that antigen B (AgB) possesses the highest diagnostic value among these antigens. In the present work we further dissected its antigenicity by analyzing the reactivity of the same panel of sera against a set of synthetic peptides spanning the sequence of both AgB subunits. The N-terminal extension of these subunits appeared to be immunodominant in human infections. A 38-mer peptide (p176) delineated from the N-terminal extension of the AgB/1 subunit performed in an enzyme-linked immunosorbent assay with a higher diagnostic sensitivity (80%) and specificity (94%) than native AgB, Ag5, or any other peptide antigen tested against this collection of serum samples. In view of its high diagnostic value and its nature as a well-defined reproducible antigen, p176 could conveniently be used as a reference standard antigen in the diagnosis of hydatid disease.

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.

scholarly journals Complete Genome Sequence of the Telford Type S Strain of Mycobacterium avium subsp. paratuberculosis

2019 ◽  
Vol 8 (11) ◽  
Author(s):  
Rudiger Brauning ◽  
Karren Plain ◽  
Milan Gautam ◽  
Tonia Russell ◽  
C. Carolina Correa ◽  
...  
Keyword(s):  
Genome Sequence ◽  
Mycobacterium Avium ◽  
Causative Agent ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Content Type ◽  
Mycobacterium Avium Subsp Paratuberculosis

Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne’s disease (JD).

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