Comparison of culture, ELISA and LAMP-PCR for diagnosis of Mycobacterium avium subsp. Paratuberculosis

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Research Paper 01/03/2015
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Comparison of culture, ELISA and LAMP-PCR for diagnosis of Mycobacterium avium subsp. Paratuberculosis

Orkideh Heidarnejhad, Shahabeddin Safi
J. Bio. Env. Sci.6( 3), 279-288, March 2015.
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Abstract

Mycobacterium avium subsp. Paratuberculosis is the etiologic agent of Johne’s disease in the cattle. JD is a disease with considerable economic impact on dairy cattle. It is prevalent worldwide, especially in developing countries including Iran. Rapid and accurate diagnosis of the infection is essential for controlling the disease. The present study compared fecal culture, ELISA and loop-mediated isothermal amplification (LAMP) for the diagnosis of MAP in dairy cattle. A total of 225 serum and fecal samples were collected from 14 dairy cattle farms in Tehran, Iran. The fecal samples were cultured in Harrold’s egg yolk agar with and without mycobactin J. DNA was extracted directly from fecal samples, too. Serum ELISA and LAMP were compared with fecal culture in terms of sensitivity, specificity, positive predictive value, negative predictive value, likelihood ratio of a positive test result, and likelihood ratio of a negative test result. The sensitivity of LAMP and ELISA was 100% and 79%, respectively. Their specificity was 83.33% and 93%, respectively. Both LAMP (P < 0.001; k = 0.167) and ELISA (P < 0.001; k = 0.374) were in agreement with fecal culture as the gold standard. Moreover, LAMP and ELISA had 80% agreement with each other. This study suggests LAMP as a valuable and cost-effective tool for the early diagnosis of JD caused by MAP. Furthermore, due to inability of ELISA in detecting early stages of the disease and long and expensive method of culture, LAMP could be used alongside ELISA in cattle-wide screening for disease.

VIEWS 8

Bull TJ, Hermon-Taylor J, Pavlik I, I, El-Zaatari F, Tizard M. 2000. Characterization of IS900 loci in mycobacterium avium subsp. paratuberculosis and development of multiplex PCR typing. Microbiology . 146 (12), 3285.

Carvalho IA, Campos VEB, Souza IM, Zanardo LG, Filho JDR, Gomes MJP, et al. 2012. Diagnosis of paratuberculosis in cattle: microbiological culture, serology and PCR. Brazil Journal of Microbiology.; 43(2), 581–585

Castellanos E, de JL, Dominguez L, Aranaz A. 2012. Progress in molecular typing of Mycobacterium avium subspecies paratuberculosis. Res Vet Sci. Apr.92(2), 169 -79.

Chamberlin W, Graham DY, Hulten K, El-Zimaity HM, Schwartz MR, Naser S, et al. 2001. Review article: Mycobacterium avium subsp. paratuberculosis  as  one  cause  of  Crohn’s  disease. Alimentary Pharmacologic Therapy. 15(3), 337-46.

Clark DL Jr, Koziczkowski JJ, Radcliff RP, Carlson RA, Ellingson JL. 2008. Detection of Mycobacterium avium subspecies paratuberculosis: comparing fecal culture versus serum enzyme-linked immunosorbent assay and direct fecal polymerase chain reaction. J Dairy Sci. 91(7), 2620-7

Collins MT.  1996.  Diagnosis  of  Paratuberculosis. Veterinary Clinical North American Food Animal Practice. 12, 357-371

Khare S, FichtT A, Santos RL, Romano J, Ficht AR, Zhang S, et al. 2004. 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. Journal of Clinical Microbiology. 42(3), 1075–1081.

Melville L, Kenyon F, Javed S, McElarney I, Demeler J, Skuce P. 2014. Development of a loop-mediated isothermal amplification (LAMP) assay for the sensitive detection of Haemonchus contortus eggs in ovine fecal samples. Veterinary Parasitology. 206, 308–312

Mori Y, Notomi T. 2009. Loop-mediated isothermal amplification (LAMP): a rapid,accurate, and cost-effective diagnostic method for infectious diseases. Journal of Infection Chemotherapy. 15(2), 62-9.

Möbius P, Hotzel H, Rassbach A, Köhler H. 2008. Comparison of 13 single-round and nested PCR assays targeting IS900, ISMav2, f57 and locus 255 for detection of Mycobacterium avium subsp. paratuberculosis. Veterinary Microbiology. 25(4), 324-33

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, et al. 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acid Reserch. 15; 28(12), E63.

OIE Terrestrial Manual. 2014. Chapter 2.1.11. Paratuberculosis (Johne’sdisease)

Parida M, Sannarangaiah S, Dash PK, Rao PV, Morita K. 2008. Loop mediated isotherma-lamplification (LAMP): a new generation of innovative gene amplification technique; perspectives in clinical diagnosis of infectious diseases. Rev Med Virol. 18(6), 407-21

Pinedo PJ, Rae DO, Williams JE, Donovan GA, Melendez P, Buergelt CD. 2008. Association among results of serum ELISA, faecal culture and nested PCR on milk, blood and faeces for the detection of paratuberculosis in dairy cows. Transboundary and emerging diseases. 55 (2), 125-133

Safi S, Heidarnejhad O, Mosavari N, Sakha M, Afshar D, Moazami L, et al. 2014.Comparative evaluation of LAMP and Nested-PCR for the diagnosis of bovine paratuberculosis. International Journal of Mycobacteriology, In Press, Corrected Proof, Available online 30 October

Saleh  M,  Soliman  H,  El-Matbouli  M.  2008. Loop-mediated isothermal amplification as anemerging technology for detection of Yersinia ruckeri the causative agent of enteric red mouth disease in fish. BMC Veterinary Reserch. 4, 31.

Salgado M, Steuer P, Troncoso E, Collins MT. 2013. Evaluation of PMS-PCR technology for detection of Mycobacterium avium subsp. paratuberculosis directly from bovine fecal specimens. Veterinary Microbiology. 27; 167(3-4), 725-8

Sohal JS, Singh SV, Subhodh S, Singh AV, Singh PK, Sheoran N, et al. 2007. Mycobacterium avium subspecies paratuberculosis diagnosis and strain typing present status and future developments. Indian Journal of Experimental Biology . 45(10), 843-52.

Tiwari A, Vanleeuwen JA, McKenna SL, Keefe GP, Barkema HW. 2006. Johne’s disease in Canada Part I: clinical symptoms, pathophysiology, diagnosis, and prevalence in dairy herds. Canadian Veterinary Journal. 47(9), 874-82.

Van Soolingen DV. 2001. Molecular epidemiology of tuberculosis and other mycobacterial infections: main methodologies and achievements. Journal of Internal Medicine: 249, 1-26.

Whitlock RH, Wells SJ, Sweeney RW, Van Tiem J. 2010. ELISA and fecal culture for paratuberculosis (Johne’s disease): sensitivity and specificity of each method. Veterinary Microbiology. 77(3-4), 387-98.

Whittington RJ. 2009. Factors affecting isolation and  identification  of  Mycobacteriumavium  subsp. paratuberculosis from fecal and tissue samples in a liquid culturesystem. Journal of Clinical Microbiology. 47(3), 614-22.

Yang W, Ying F, Yingyu L, Pin C, Wentao L, Shuqing  L,  et  al.   2010.   Development   and evaluation of loop-mediated isothermal amplification for rapid detection of Haemophilus parasuis. FEMS Microbiology Letter.313(1), 54-60.