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The First Prevalence Report of Direct Identification and Differentiation of B. abortus and B. melitensis using Real Time PCR in House Mouse of Iran

Authors: A. Doosti, S. Moshkelani


Brucellosis is a zoonotic disease; its symptoms and appearances are not exclusive in human and its traditional diagnosis is based on culture, serological methods and conventional PCR. For more sensitive, specific detection and differentiation of Brucella spp., the real time PCR method is recommended. This research has performed to determine the presence and prevalence of Brucella spp. and differentiation of Brucella abortus and Brucella melitensis in house mouse (Mus musculus) in west of Iran. A TaqMan analysis and single-step PCR was carried out in total 326 DNA of Mouse's spleen samples. From the total number of 326 samples, 128 (39.27%) gave positive results for Brucella spp. by conventional PCR, also 65 and 32 out of the 128 specimens were positive for B. melitensis, B. abortus, respectively. These results indicate a high presence of this pathogen in this area and that real time PCR is considerably faster than current standard methods for identification and differentiation of Brucella species. To our knowledge, this study is the first prevalence report of direct identification and differentiation of B. abortus and B. melitensis by real time PCR in mouse tissue samples in Iran.

Keywords: Differentiation, Iran, B. abortus, B. melitensis, TaqManprobe

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[1] A. Al-Mariri, P. A. Tibor, X. Mertens, P. De Bolle, J. Michel, G. K. Walravens, and J. J. Letesson, "Induction of immune response in BALB/c mice with a DNA vaccine encoding bacterioferritin or P39 of Brucella spp.," Infect. Immun., vol. 69, pp. 6264-6270, Apr. 2001.
[2] W. S. Probert, K. N. Schrader, N. Y. Khuong, S. L. Bystrom and M. H. Graves, "Real-Time multiplex PCR assay for detection of Brucella spp., B. abortus, and B. melitensis," J. Clin. Microbiol., 2004; vol. 42, no. 3, pp. 1290-1293, Jan. 2004.
[3] B. Kazemi, S. A. Yousefi Namin, M. Dowlatshahi, M. Bandepour, F. Kafilzadeh, L. Gachkar, F. Mahmoudinejad, A. Samarghandi, and M. Mardani, "Detection of Brucella by peripheral blood PCR and comparison with culture and serological methods in suspected cases." Iranian J. Publ. Health., vol. 37, no. 4, pp. 96-102, Agu. 2008.
[4] T. Bogdanovich, M. Skurnik, P. S. Lubeck, P. Ahrens, and J. Hoorfar, "Validated 5' nuclease PCR assay for rapid identification of the genus Brucella." J. Clin. Microbiol., vol. 42, no. 5, pp. 2261-2263, Des. 2004.
[5] B. J. Bricker, "PCR as a diagnostic tool for brucellosis." Vet. Microbiol., vol. 90, pp. 435-446, Jun. 2002.
[6] C. Romero, C. Gamazo, M. Pardo, and I. Lopez-Goni, "Specific detection of Brucella DNA by PCR." J. Clin. Microbiol., vol. 33, pp. 615-617, Jul. 1995.
[7] T. J. Carver, K. M. Rutherford, M. Berriman, M. A. Rajandream, B. G. Barrell, and J. Parkhill, ACT: the Artemis Comparison Tool." Bioinformatics., vol. 21, pp. 3422-3423, Aug. 2005.
[8] E. Navarro, M. A. Casao, and J. Solera, "Diagnosis of human brucellosis using PCR." Expert Rev. Mol. Diagn., vol. 4, pp. 115-123, Oct. 2004.
[9] B. J. Bricker, and S. M. Halling, "Differentiation of Brucella abortus bv 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv 1 by PCR" J. Clin. Microbiol., vol. 32, pp. 2660-2666, Feb. 1994.
[10] B. J. Bricker, "Differentiation of hard-to-type bacterial strains by RNA mismatches cleavage." Biotechniques., vol. 27, pp. 321-324, Jul. 1999.
[11] E. Tcherneva, N. Rijpens, B. Jersek, and L. M. Herman, "Differentiation of Brucella species by random amplified polymorphic DNA analysis. Journal of Applied Microbiol., vol. 88, pp. 69-80, Aug. 2000.
[12] J. T. Foster, R. T. Okinaka, R. Svensson, K. Shaw, B. K. De, A. R. Robison, W. S. Probert, L. J. Kenefic, W. D. Brown, and P. Keim, "Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades." J. Clin. Microbiol., vol. 46, pp. 296-301, Jan. 2008.
[13] J. Sambrook, D. W. Russell, Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York: 2001, 3rd edn.
[14] D. Leal-Klevezas, I.O. Martinez vazquez, A. Lopez-Merino, Martinez- J. P. Soriano, "Single-step PCR for detection of Brucella spp. from blood and milk of infected animals." J. Clin. Microbiol., vol. 33, no.12, pp. 3087-3090, Des. 1995.
[15] B. Osterman, and I. Moriyo'n, "International Committee on Systematics of Prokaryotes; subcommittee on the taxonomy of Brucella: minutes of the meeting, 17 September 2003, Pamplona, Spain." Int. J. Syst. Evol. Microbiol., vol. 56, pp. 1173-1175, Jun. 2006.
[16] B. G. Mantur, S. K. Amarnath, and R. S. Shinde "Review of clinical and laboratory features of human brucellosis." Indian journal of medical microbiology., vol. 25, no. 3, pp. 188-202, Des. 2007.
[17] Y. A. Al-Eissa, "Brucellosis in Saudi Arabia: past, present and future." Annals of Saudi medicine., vol. 19, no. 5, 401-405, May. 1999.
[18] P. A. Vershilova, G. Liamkin, V. E. Malikov, E. A. Dranovskaya, and I. F. Taran, "Brucella strains from Mouselike Rodents in Southwestern USSR." International Jolu Rnal of Systematbaicc Teriology., vol. 33, no. 2, pp. 399-400, Mar. 1983.
[19] C. Romero, C. Gamazo, M. Pardo, and I. Lopez-Goni, "Specific Detection of Brucella DNA by PCR." J. Clin. Microbiol., vol. 33, no.3, pp. 615-617, Mar. 1995.
[20] E. Navarro, M. A. Casao, and J. Solera, "Diagnosis of human brucellosis using PCR." Expert Rev. Mol. Diagn., vol. 4, no.1, pp. 115-123, Feb. 2004.
[21] M. J. LaGier, L. A. Joseph, T. V. Passaretti, K. A. Musser, and N. M. Cirino, "A real-time multiplexed PCR assay for rapid detection and differentiation of Campylobacter jejuni and Campylobacter coli." Molecular and Cellular Probes., vol. 18, pp. 275-282, Jul. 2004.
[22] L. Bounaadja, D. Albert, B. Che nais, S. Henault, M.S. Zygmunt, S. Poliak, and B. Garin-Bastuji, "Real-time PCR for identification of Brucella spp.: A comparative study of IS711, bcsp31 and per target genes." Vet. Microbiol., vol. 137, pp. 156-164, Nov. 2009.