Development of a bead-based Luminex assay using lipopolysaccharide specific monoclonal antibodies to detect biological threats from Brucella species

Table 2 Specificity of the developed bead-based Luminex immunoassay

Bacterial species	Strain	O-Antigen	Luminex assay (mean fluorescence intensity)
Brucella spp. (biotype)
B. abortus (1)	NCTC 10093 544	A^1,3	13107
	19S	A^1,3	11300
	A146-10	A^1,3	5607
B. abortus (3)	A104-10 Uckermark	A^1,3	3172
B. canis	NCTC 10854 RM-666	R^2,3	33
B. ovis	CNCTC 6741	R^2,3	23
B. melitensis (1)	NCTC 10094 16 M	M^1,3	25860
	ICM 3.33	M^1,3	13129
	ICM 583/2003	M^1,3	16972
	ICM 91/2004	M^1,3	15030
	102A01C2F	M^1,3	22285
	A146-13	M^1,3	11407
	A104-11 Tgb. Nr. 117518	M^1,3	5977
B. melitensis (2)	A104-12 799/97, B3898	A^1,3	2958
B. melitensis (3)	A104-13 210739, Mainz	AM^1,3	1884
B. suis (1)	NCTC 10316 1330	A^1,3	5167
B. suis (2)	A 104–14 Rostock	A^1,3	10316
B. neotomae	A148-7 5 K33	A^1,3	1875
Other bacteria
F. tularensis tularensis	ATCC 6223	D⁴	18
F. tularensis holarctica	NCTC 10857	D⁴	22
B. anthracis (spores)	Böhm 73202.2000 (PX02)	D⁵	19
S. typhimurium	ATCC 14028	D⁶	19
Y. enterocolitica	310 (IT2, ST9) O9	A^7,8,9	1291
Y. pestis	CO92	R¹⁰	20
O. anthropi	ATCC 49188	D¹¹	20
E. coli	O157, 15326	A^7,12	22
B. mallei	NCTC 03709 106	D¹³	18
B. pseudomallei	H05410-0490	D¹⁴	20
V. cholera	O1, ATCC 14734	A¹⁵	24

Luminex LOD was defined as two times the mean fluorescence intensity of the blank (mean blank = 20) and used as the threshold for positive results. Values in bold indicate positive results. Classification of O-antigens [4, 26–29, 41–50]: A = α1,2-linked 4-amino-4,6-dideoxy-α-D-mannopyranosyl subunits, M = α1,3-linked and α1,2-linked 4,6-dideoxy-4-formamido-α-D-mannopyranosyl residues, D = different O-antigen structure compared to Brucella, R = ‘rough’ LPS (no O-antigen). Meikle et al. 1989¹, Adone et al. 2011², Corbel 2006³, Wang et al. 2011⁴, Crich and Vinogradova 2007⁵, Watson et al. 1992⁶, Perry et al. 1986⁷, Caroff et al. 1984⁸, Bundle et al. 1984⁹, Skurnik et al. 2000¹⁰, Velasco et al. 1996¹¹, Perry and Bundle 1990¹², Burtnick et al. 2002¹³, Perry et al. 1995¹⁴, Kenne et al. 1982¹⁵