Rapid identification of Brucella isolates to the species level by real time PCR based single nucleotide polymorphism (SNP) analysis

Table 1 Gene targets, primers and allele-specific probes used in this study.

Species	Gene target	Location in B. abortus 9–941	Primers		Probes
B. abortus	fba A	AE017224	F	TGACATCATGCTCCGTCACATG	VIC	ATGCCGTGG CGGAA
		360225–361289	R	CAGACCGGAATATGCGGATAGAT	FAM	ATGCCGTGA CGGAA
B. melitensis	gap	AE017223	F	GGCTCAGGTTCTCAACGATACTATC	VIC	CGTGGTCATA AAGC
		1684721–1685728	R	TCGCCCGTATAGGAGTGGAT	FAM	CGTGGTCATG AAGC
B. ovis	aro A	AE017223	F	CGACCACCGCATCGC	VIC	CCATGACA AGGAAAC
		29246–30598	R	CCGGCTTTTCCGATGCAA	FAM	CATGACG AGGAAAC
B. suis bv1–4	prpE	AE017223	F	GCGACCGCATCCTCATCTATATG	VIC	CAAGCG TGGCAACC
		1687718–1689625	R	CGCCGAATACGACGGAATGAAT	FAM	CAAGCA TGGCAACC
Marine mammal Brucellae	trp E	AE017223	F	CGAGGATTCCTTCGTCCATACG	VIC	CCAATTATTTCCA CCAGACG
		1537355–1539550	R	ACGCACGGTGGAAACCTT	FAM	CCAATTATTTCCG CCAGACG
B. canis	omp 25	AE017223	F	GCTGGCGCCTTTGCT	VIC	AACTTCCAGA AGGACC
		710024–710625	R	GGCCGTCCTTGGACTTCTTG	FAM	AACTTCCAGC AGGACC
B. neotomae	putative oxido-reductase	AE017223	F	GGTTTTCCATGCGGTTTATTTGC	VIC	CATTGAGT GGCCCGAT
		1989869–1990870	R	GGCATCATGCACAGTGATATCGA	FAM	ATTGAGC GGCCCGAT
16SrRNA_771/778	16SrRNA	-	F	CGCCGTAAACGATGAATGTTAGC	VIC	CGAAGT GTAAACA CCCCGA
			R	GCGGAATGTTTAATGCGTTAGCT	FAM	CGAAGA GTAAACT CCCCGA
16SrRNA₁₀₅₅	16SrRNA	-	F	GGGTTAAGTCCCGCAACGA	VIC	ACCCTCGCCT TTAGTT
			R	ACGTCATCCCCACCTTCCT	FAM	CCCTCGCCC TTAGTT

Gene targets, primers and allele-specific probes used in this study. Brucella species-specific SNPs and non-specific SNPs (both underlined) are shown within the relevant probes. Gene target locations are given relative to the sequenced B. abortus 9-941 strain.

ISSN: 1471-2180