Comparison of Real-Time, Quantitative PCR with Molecular Beacons to Nested PCR and Culture Methods for Detection of Mycobacterium avium subsp. paratuberculosis in Bovine Fecal Samples

ABSTRACT An automated PCR with fluorescent probes (molecular beacons) detected Mycobacterium avium subsp. paratuberculosis in bovine feces. When the PCR was compared with culture in testing 41 fecal samples, kappa scores of 0.94 to 0.96, a sensitivity of 93 to 96%, and a specificity of 92% were obtained. Results were quantitated by using a standard curve derived from a plasmid containing IS900. A minimum quantity of 1.7 × 10−4 pg of DNA, correlating to 1 to 8 CFU, was detected.

[1]  A. Larsen,et al.  Age of cattle as related to resistance to infection with Mycobacterium paratuberculosis. , 1975, American journal of veterinary research.

[2]  K. White,et al.  Clinical epidemiology. , 1983, International journal of epidemiology.

[3]  H. V. Van Kruiningen,et al.  Ruminant paratuberculosis (Johne's disease): the current status and future prospects. , 1984, The Cornell veterinarian.

[4]  W. Burnham.  [Role of mycobacteria in Crohn's disease]. , 1989, Gastroenterologie clinique et biologique.

[5]  T. Picknett,et al.  Transformation of Penicillium chrysogenum with selection for increased resistance to benomyl. , 1989, FEMS microbiology letters.

[6]  R. Chiodini,et al.  Crohn's disease and the mycobacterioses: a review and comparison of two disease entities , 1989, Clinical Microbiology Reviews.

[7]  J. McFadden,et al.  Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis. , 1989, Nucleic acids research.

[8]  D. Gabrić,et al.  Identification of a repetitive DNA sequence specific to Mycobacterium paratuberculosis. , 1989, FEMS microbiology letters.

[9]  J. McFadden,et al.  Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease , 1990, Journal of clinical microbiology.

[10]  R. Yolken,et al.  Removal of inhibitory substances from human fecal specimens for detection of group A rotaviruses by reverse transcriptase and polymerase chain reactions , 1990, Journal of clinical microbiology.

[11]  D. Callihan,et al.  Cultivation of Mycobacterium Paratuberculosis from Bovine Fecal Specimens and a Suggested Standardized Procedure , 1991, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[12]  P. Kapke,et al.  Comparison of a Commercial DNA Probe Test and Three Cultivation Procedures for Detection of Mycobacterium Paratuberculosis in Bovine Feces , 1992, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[13]  R. Sweeney,et al.  Isolation of Mycobacterium paratuberculosis after oral inoculation in uninfected cattle. , 1992, American journal of veterinary research.

[14]  R. Haring,et al.  Evaluation of the abilities of three diagnostic tests based on the polymerase chain reaction to detect Mycobacterium paratuberculosis in cattle: application in a control program , 1992, Journal of clinical microbiology.

[15]  D. Collins,et al.  Comparison of polymerase chain reaction tests and faecal culture for detecting Mycobacterium paratuberculosis in bovine faeces. , 1993, Veterinary microbiology.

[16]  N. Brousse,et al.  Detection of Mycobacterium paratuberculosis by polymerase chain reaction in children with Crohn's disease. , 1994, The Journal of infectious diseases.

[17]  Sanjay Tyagi,et al.  Molecular Beacons: Probes that Fluoresce upon Hybridization , 1996, Nature Biotechnology.

[18]  G. Baljer,et al.  Molecular characterization of Mycobacterium paratuberculosis isolates from sheep, goats, and cattle by hybridization with a DNA probe to insertion element IS900 , 1996, Journal of clinical microbiology.

[19]  M. Collins.  Diagnosis of paratuberculosis. , 1996, The Veterinary clinics of North America. Food animal practice.

[20]  M. Sunairi,et al.  Rapid and efficient method to eliminate substances inhibitory to the polymerase chain reaction from animal fecal samples. , 1996, Veterinary microbiology.

[21]  C. Rossiter,et al.  Farm-specific approach to paratuberculosis (Johne's disease) control. , 1996, The Veterinary clinics of North America. Food animal practice.

[22]  J. Stabel.  An Improved Method for Cultivation of Mycobacterium Paratuberculosis from Bovine Fecal Samples and Comparison to Three Other Methods , 1997, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[23]  A. Pell,et al.  Molecular beacons: trial of a fluorescence-based solution hybridization technique for ecological studies with ruminal bacteria , 1997, Applied and environmental microbiology.

[24]  Fred Russell Kramer,et al.  Multicolor molecular beacons for allele discrimination , 1998, Nature Biotechnology.

[25]  Amalio Telenti,et al.  Molecular beacon sequence analysis for detecting drug resistance in Mycobacterium tuberculosis , 1998, Nature Biotechnology.

[26]  H. Blom,et al.  Molecular beacons: a new approach for semiautomated mutation analysis. , 1998, Clinical chemistry.

[27]  M. D. Perkins,et al.  Comparison of the ABI 7700 System (TaqMan) and Competitive PCR for Quantification of IS6110 DNA in Sputum during Treatment of Tuberculosis , 1998, Journal of Clinical Microbiology.

[28]  Fred Russell Kramer,et al.  Spectral Genotyping of Human Alleles , 1998, Science.

[29]  F. Rommel,et al.  Mycobacterium Paratuberculosis Detection in Bovine Feces is Improved by Coupling Agar Culture Enrichment to an IS900-Specific Polymerase Chain Reaction Assay , 1999, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc.

[30]  D. Ho,et al.  Use of Real-Time PCR and Molecular Beacons To Detect Virus Replication in Human Immunodeficiency Virus Type 1-Infected Individuals on Prolonged Effective Antiretroviral Therapy , 1999, Journal of Virology.

[31]  Sanjay Tyagi,et al.  Multiplex detection of four pathogenic retroviruses using molecular beacons. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[32]  F. Veglia,et al.  Real-Time Quantitative PCR for Human Herpesvirus 6 DNA , 2000, Journal of Clinical Microbiology.

[33]  M. Drake,et al.  Molecular beacon polymerase chain reaction detection of Escherichia coli O157:H7 in milk. , 2000, Journal of food protection.

[34]  M. Wong,et al.  Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon , 2000, Journal of Clinical Microbiology.

[35]  S. Wong,et al.  Molecular beacons: a new approach to plant virus detection. , 2000, Phytopathology.

[36]  John P. Moore,et al.  Global distribution of the CCR2-64I/CCR5-59653T HIV-1 disease-protective haplotype , 2000, AIDS.

[37]  N. Nowak,et al.  Semiautomated clone verification by real-time PCR using molecular beacons. , 2000, BioTechniques.

[38]  N. Høiby,et al.  Results of Multiple Diagnostic Tests forMycobacterium avium subsp. paratuberculosis in Patients with Inflammatory Bowel Disease and in Controls , 2000, Journal of Clinical Microbiology.

[39]  P. Rådström,et al.  Effects of Amplification Facilitators on Diagnostic PCR in the Presence of Blood, Feces, and Meat , 2000, Journal of Clinical Microbiology.

[40]  A. Syvänen,et al.  Homogeneous scoring of single-nucleotide polymorphisms: comparison of the 5'-nuclease TaqMan assay and Molecular Beacon probes. , 2000, BioTechniques.

[41]  M. Washington,et al.  Johne's disease in cattle , 1990, The Bovine Practitioner.

[42]  D. Cousins.  Paratuberculosis Johne ’ s Disease , 2019 .

[43]  R. Henry,et al.  Detection of Transgenes in Crop Plants Using Molecular Beacon Assays , 2004, Plant Molecular Biology Reporter.

[44]  R. Smith,et al.  Veterinary Clinics of North America: Food Animal Practice , 2019, Veterinary Clinics of North America: Food Animal Practice.