Detection of Mycobacterium avium subsp. paratuberculosis by a Sonicate Immunoassay Based on Surface-Enhanced Raman Scattering

ABSTRACT A sandwich immunoassay for the rapid, low-level detection of Mycobacterium avium subsp. paratuberculosis has been developed. M. avium subsp. paratuberculosis is the causative agent of Johne's disease in cattle, and one of the major obstacles in controlling the spread of this disease is the inability to rapidly detect small amounts of bacteria or other diagnostic markers shed during the subclinical stage of infection. This paper details the development and performance of an assay for sonicated M. avium subsp. paratuberculosis lysate that is based on surface-enhanced Raman scattering (SERS). There are two key components of the assay: (i) an immobilized layer of monoclonal antibodies that target a surface protein on the microorganism; and (ii) extrinsic Raman labels (ERLs) that are designed to selectively bind to captured proteins and produce large SERS signals. By correlating the number of M. avium subsp. paratuberculosis bacilli present prior to sonication to the amount of total protein in the resulting sonicate, the detection limit determined for total protein can be translated to the microorganism concentration. These findings yield detection limits of 100 and 200 ng/ml (estimated to be 500 and 1,000 M. avium subsp. paratuberculosis bacilli/ml) for sonicate spiked in phosphate buffer and sonicate spiked in whole milk, respectively. Moreover, the time required to complete the assay, which includes sample preparation, antigen extraction, ERL incubation, and readout, is less than 24 h. The potential for incorporation of this novel assay into diagnostic laboratories is also briefly discussed.

[1]  W. Bishai,et al.  Evaluation of the accuracy and reproducibility of a practical PCR panel assay for rapid detection and differentiation of Mycobacterium avium subspecies. , 2000, Molecular and cellular probes.

[2]  T. Cotton,et al.  Immunoassay employing surface-enhanced Raman spectroscopy. , 1989, Analytical biochemistry.

[3]  B. Narasimhan,et al.  Combinatorial materials science , 2007 .

[4]  V. Kapur,et al.  Genome Scale Comparison of Mycobacterium avium subsp. paratuberculosis with Mycobacterium avium subsp. avium Reveals Potential Diagnostic Sequences , 2002, Journal of Clinical Microbiology.

[5]  J. Gala,et al.  Definitive Differentiation between Single and Mixed Mycobacterial Infections in Red Deer (Cervus elaphus) by a Combination of Duplex Amplification of p34 and f57 Sequences and Hpy188I Enzymatic Restriction of Duplex Amplicons , 2005, Journal of Clinical Microbiology.

[6]  S. Sreevatsan,et al.  Development and Characterization of Monoclonal Antibodies and Aptamers against Major Antigens of Mycobacterium avium subsp. paratuberculosis , 2007, Clinical and Vaccine Immunology.

[7]  L. V. van Keulen,et al.  Paratuberculosis in sheep: its possible role in the epidemiology of paratuberculosis in cattle. , 2001, Veterinary microbiology.

[8]  Olga Lyandres,et al.  Rapid detection of an anthrax biomarker by surface-enhanced Raman spectroscopy. , 2005, Journal of the American Chemical Society.

[9]  J. Popp,et al.  Surface-enhanced Raman spectroscopy , 2009, Analytical and bioanalytical chemistry.

[10]  J. M. Sharp,et al.  Paratuberculosis Infection of Nonruminant Wildlife in Scotland , 2001, Journal of Clinical Microbiology.

[11]  M. Porter,et al.  Femtomolar detection of prostate-specific antigen: an immunoassay based on surface-enhanced Raman scattering and immunogold labels. , 2003, Analytical chemistry.

[12]  M. Collins,et al.  Evaluation of four serological tests for bovine paratuberculosis , 1992, Journal of clinical microbiology.

[13]  George M. Whitesides,et al.  Features of gold having micrometer to centimeter dimensions can be formed through a combination of stamping with an elastomeric stamp and an alkanethiol ‘‘ink’’ followed by chemical etching , 1993 .

[14]  R W Sweeney,et al.  Transmission of paratuberculosis. , 1994, The Veterinary clinics of North America. Food animal practice.

[15]  Marc D. Porter,et al.  Designing Interfaces at the Molecular Level , 1995 .

[16]  P. Beard,et al.  Natural paratuberculosis infection in rabbits in Scotland. , 2001, Journal of comparative pathology.

[17]  S. More,et al.  Sensitivity and specificity of pooled faecal culture and serology as flock-screening tests for detection of ovine paratuberculosis in Australia. , 2002, Preventive veterinary medicine.

[18]  J. Griffin,et al.  Mycobacterial diseases of deer , 2004, New Zealand veterinary journal.

[19]  I. Kyriazakis,et al.  Clustering of Mycobacterium avium subsp. paratuberculosis in Rabbits and the Environment: How Hot Is a Hot Spot? , 2005, Applied and Environmental Microbiology.

[20]  Robert J. Lipert,et al.  Impact of Protein Shedding on Detection of Mycobacterium avium subsp. paratuberculosis by a Whole-Cell Immunoassay Incorporating Surface-Enhanced Raman Scattering , 2007, Clinical and Vaccine Immunology.

[21]  S. Wells,et al.  Herd-level economic losses associated with Johne's disease on US dairy operations. , 1999, Preventive veterinary medicine.

[22]  R. Dasari,et al.  Surface-enhanced Raman scattering and biophysics , 2001 .

[23]  D. Hunter,et al.  Rapid and Sensitive Detection of Mycobacterium avium subsp. paratuberculosis in Bovine Milk and Feces by a Combination of Immunomagnetic Bead Separation-Conventional PCR and Real-Time PCR , 2004, Journal of Clinical Microbiology.

[24]  J. Stabel,et al.  Evaluation of a commercial ELISA for diagnosis of paratuberculosis in cattle. , 2001, Journal of the American Veterinary Medical Association.

[25]  M. Porter,et al.  Surface-enhanced Raman scattering immunoassays using a rotated capture substrate. , 2007, Analytical chemistry.

[26]  J. Miller,et al.  Early Induction of Humoral and Cellular Immune Responses during Experimental Mycobacterium avium subsp. paratuberculosis Infection of Calves , 2003, Infection and Immunity.

[27]  M. Porter,et al.  Ultrasensitive Immunoassays Based on Surface-Enhanced Raman Scattering by Immunogold Labels , 2006 .

[28]  M. Porter,et al.  Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids. , 1999, Analytical chemistry.

[29]  R. Evans,et al.  Mycobacteria distenct from Mycobacterium avium subsp. paratuberculosis isolated from the faeces of ruminants possess IS900-like sequences detectable IS900 polymerase chain reaction: implications for diagnosis. , 1999, Molecular and cellular probes.

[30]  J. Álvarez,et al.  Paratuberculosis and avian tuberculosis infections in one red deer farm studied by IS900 and IS901 RFLP analysis. , 2005, Veterinary microbiology.

[31]  R. Whittington,et al.  Intrauterine and transmammary transmission of Mycobacterium avium subsp paratuberculosis in sheep. , 2004, Australian veterinary journal.

[32]  Stephen L. Ott,et al.  JOHNE'S DISEASE ON U.S. DAIRY OPERATIONS , 1997 .

[33]  A. Ulman,et al.  Formation and Structure of Self-Assembled Monolayers. , 1996, Chemical reviews.

[34]  Shuming Nie,et al.  Surface-enhanced Raman spectroscopic detection of cancer biomarkers in intact cellular specimens , 2005, SPIE BiOS.

[35]  H. Brun-Hansen,et al.  Subclinical paratuberculosis in goats following experimental infection. An immunological and microbiological study. , 2001, Veterinary immunology and immunopathology.

[36]  Robert J. Lipert,et al.  Single-particle Raman measurements of gold nanoparticles used in surface-enhanced Raman scattering (SERS)-based sandwich immunoassays , 2004, SPIE Optics East.

[37]  I. Grant,et al.  Incidence of Mycobacterium paratuberculosis in Bulk Raw and Commercially Pasteurized Cows' Milk from Approved Dairy Processing Establishments in the United Kingdom , 2002, Applied and Environmental Microbiology.

[38]  L. Bermudez,et al.  Johne's disease, inflammatory bowel disease, and Mycobacterium paratuberculosis. , 2004, Annual review of microbiology.

[39]  James E. Collins,et al.  Evaluation of Five Antibody Detection Tests for Diagnosis of Bovine Paratuberculosis , 2005, Clinical Diagnostic Laboratory Immunology.

[40]  Duncan Graham,et al.  Simple multiplex genotyping by surface-enhanced resonance Raman scattering. , 2002, Analytical chemistry.

[41]  Marc D Porter,et al.  Labeled gold nanoparticles immobilized at smooth metallic substrates: systematic investigation of surface plasmon resonance and surface-enhanced Raman scattering. , 2006, The journal of physical chemistry. B.

[42]  Robert D. Herbert,et al.  What do I need to know , 2005 .

[43]  L. Bermudez,et al.  The Mycobacterium avium subsp. paratuberculosis 35 kDa protein plays a role in invasion of bovine epithelial cells. , 2003, Microbiology.

[44]  R W Sweeney,et al.  ELISA and fecal culture for paratuberculosis (Johne's disease): sensitivity and specificity of each method. , 2000, Veterinary microbiology.

[45]  J. C. Low,et al.  Comparison of the absorbed ELISA and agar gel immunodiffusion test with clinicopathological findings in ovine clinical paratuberculosis , 1996, Veterinary Record.

[46]  J. Stabel,et al.  Johne's disease: a hidden threat. , 1998, Journal of dairy science.

[47]  R. Whitlock,et al.  Typing of IS 1311 polymorphisms confirms that bison (Bison bison) with paratuberculosis in Montana are infected with a strain of Mycobacterium avium subsp. paratuberculosis distinct from that occurring in cattle and other domesticated livestock. , 2001, Molecular and cellular probes.

[48]  M. Natan,et al.  Glass-Coated, Analyte-Tagged Nanoparticles: A New Tagging System Based on Detection with Surface-Enhanced Raman Scattering , 2003 .

[49]  B. Brooks,et al.  PARATUBERCULOSIS IN SAIGA ANTELOPE (SAIGA TATARICA) AND EXPERIMENTAL TRANSMISSION TO DOMESTIC SHEEP , 1992, Journal of wildlife diseases.

[50]  J. Stabel,et al.  Production and Characterization of Monoclonal Antibodies against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis , 2007, Clinical and Vaccine Immunology.

[51]  G. Gerlach,et al.  Development of a Peptide-Mediated Capture PCR for Detection of Mycobacterium avium subsp. paratuberculosis in Milk , 2002, Journal of Clinical Microbiology.

[52]  Duncan Graham,et al.  DNA detection by surface enhanced resonance Raman scattering (SERRS). , 2005, The Analyst.

[53]  Hiroshi Yamamoto,et al.  Enzyme Immunoassay Utilizing Surface-Enhanced Raman Scattering of the Enzyme Reaction Product , 1997 .

[54]  I. Berg,et al.  Evaluation of the gamma interferon test for diagnosis of paratuberculosis in goats. , 2005, Veterinary immunology and immunopathology.

[55]  C. Shannon,et al.  Heterogeneous immunosensing using antigen and antibody monolayers on gold surfaces with electrochemical and scanning probe detection. , 2000, Analytical chemistry.

[56]  M. S. Burton,et al.  MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS INFECTION IN AN ADDAX (ADDAX NASOMACULATUS) , 2009, Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians.

[57]  Yukihiro Ozaki,et al.  Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering. , 2004, The Analyst.

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

[59]  C. Thoen,et al.  Paratuberculosis in cattle: a comparison of three serologic tests with results of fecal culture. , 1989, Veterinary microbiology.

[60]  C. Mirkin,et al.  Nanoparticles with Raman spectroscopic fingerprints for DNA and RNA detection. , 2002, Science.

[61]  M. Porter,et al.  Low-level detection of viral pathogens by a surface-enhanced Raman scattering based immunoassay. , 2005, Analytical chemistry.