Rapid Multiplex PCR Assay for the Simultaneous Detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis = Rapid Multiplex PCR Assay for the Simultaneous Detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis

다국어 초록 (Multilingual Abstract)

The routine identification and differentiation of Brucella species is a time-consuming and labor-intensive process, which frequently places personnel at risk of laboratory-acquired infection. Here, we describe the development of a rapid multiplex PCR assay for the confirmation of presumptive Brucella isolates. The assay was able to identify and differentiate major human pathogens, namely B. abortus, B. melitensis, and B. suis, in a single test of less than an hour and a half.

참고문헌 (Reference)

1 Bogdanovich, T, "Validated 5' nuclease PCR assay for rapid identification of the genus Brucella" 42 : 2261-2263, 2004

2 Paulsen, I. T, "The Brucella suis genome reveals fundamental similarities between animal and plant pathogens and symbionts" 99 : 13148-13153, 2002

3 Alton, G. G, "Techniques for Brucellosis Laboratory" INRA 1988

4 Leal-Klevezas, D. S, "Single-step PCR for detection of Brucella spp. from blood and milk of infected animals" 33 : 3087-3090, 1995

5 Singh, M, "Serum as clinical specimen in PCR for diagnosis of ovine brucellosis" 80 : 17-18, 2010

6 Probert, W. S, "Real-time multiplex PCR assay for detection of Brucella spp., B. abortus, and B. melitensis" 42 : 1290-1293, 2004

7 Redkar, R, "Real-time detection of Brucella abortus, Brucella melitensis and Brucella suis" 15 : 43-52, 2001

8 Newby, D.T, "Real-time PCR detection of Brucella abortus. A comparative study of SYBR green I, 5'-exonuclease, and hybridization probe assays" 69 : 4753-4759, 2003

9 Foster, J. T, "Real-time PCR assays of singlenucleotide polymorphisms defining the major Brucella clades" 46 : 296-301, 2008

10 Bricker, B. J, "PCR as a diagnostic tool for brucellosis" 90 : 435-446, 2002

11 Hinic, V, "Novel identification and differentiation of Brucella melitensis, B. abortus, B. suis, B. ovis, B. canis, and B. neotomae suitable for both conventional and real-time PCR systems" 75 : 375-378, 2008

12 Scott, J. C, "Multiplex assay based on single-nucleotide polymorphisms for rapid identification of Brucella isolates at the species level" 73 : 7331-7337, 2007

13 García-Yoldi, D, "Multiplex PCR assay for the identification and differentiation of all Brucella species and the vaccine strains Brucella abortus S19 and RB51 and Brucella melitensis Rev1" 52 : 779-781, 2006

14 Hoorfar, J., "Making internal amplification control mandatory for diagnostic PCR" 41 : 5835-, 2003

15 Gandara, B, "Limited genetic diversity of Brucella spp" 39 : 235-240, 2001

16 Herman, L, "Identification of Brucella spp. by using the polymerase chain reaction" 58 : 2099-2101, 1992

17 Sohn, A. H, "Human neurobrucellosis with intracerebral granuloma caused by a marine mammal Brucella spp" 9 : 485-488, 2003

18 Rijpens, N. P, "Direct detection of Brucella spp. in raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes" 62 : 1683-1688, 1996

19 Tcherneva, E, "Differentiation of Brucella species by random amplified polymorphic DNA analysis" 88 : 69-80, 2000

20 Bricker, B. J, "Differentiation of Brucella abortus bv. 1, 2 & 4, Brucella melitensis, Brucella ovis and Brucella suis bv. 1 by PCR" 32 : 2660-2666, 1994

21 Navarro, E, "Diagnosis of human brucellosis using PCR" 4 : 115-123, 2004

22 Kumar Sanjay, "Detection of Virulence-Associated Genes in Clinical Isolates of Bacillus anthracis by Multiplex PCR and DNA Probes" 한국미생물·생명공학회 19 (19): 1475-1481, 2009

23 Kumar, S, "Detection of Salmonella enterica serovar Typhi (S. typhi) by selective amplification of invA, viaB, fliC-d and prt genes by polymerase chain reaction in mutiplex format" 49 : 149-154, 2006

24 Baily, G. G, "Detection of Brucella melitensis and Brucella abortus by DNA amplification" 95 : 271-275, 1992

25 Halling, S. M, "Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis" 187 : 2715-2726, 2005

26 Rajashekara, G, "Comparative whole genome hybridization reveals genomic islands in Brucella species" 186 : 5040-5051, 2004

27 Corbel, M. J, "Brucellosis. An overview" 3 : 213-221, 1997

28 Theron, J., "A sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples" 35 : 869-874, 2001