Phenotypic and Genotypic Drug Susceptibility Assessment of Mycobacterium bovis Bacillus Calmette-Guérin Clinical Strains

Figures & data

Table 1 Clinical Characteristics of the Patients

Country	Strain	Clinical Sample	Age	Comorbidity	Year
Brazil	4BR	Lymph node	2	–	2016
	5BR	Lymph node	2	–	2016
	6BR	Blood culture	5 months	Milliary TB	2015
	7BR	Tracheal aspirate	5 months	Milliary TB	2015
	8BR	Abscess at the inoculation site	8 months	–	2015
Argentina	1AR	Lymph node	1	–	2015
	2AR	Lymph node	1	HIV+	2013
	3AR	Blood culture	–	HIV+	2005
	4AR	Bone biopsy	1	–	2007
	5AR	Gastric juice aspirate	2	–	2014
	6AR	Gastric juice aspirate	3	–	2016
	7AR	Lymph node	1	HIV+	2001
	8AR	Bone marrow	1	HIV+	2001
	9AR	Blood culture	1	HIV+	2001
	10AR	Lymph node	8	–	2003
	11AR	Lymph node	–	HIV+	2005
	12AR	Lymph node	4	HIV-	2006
	13AR	Abscess at the inoculation site	1	-	2009
	14AR	Lymph node	-	-	2012
	15AR	Lymph node	1	HIV+	2014
	16AR	Skin biopsy	-	-	2014
	17AR	Abscess at the inoculation site	2	-	2010
	18AR	Lymph node	1	HIV+	2004
	19AR	Blood culture	-	HIV+	2002
	20AR	Blood culture	-	HIV+	2003
	21AR	Blood culture	3	HIV+	2005
	22AR	Non identified	1	HIV-	2005
	23AR	Bone biopsy	1	HIV-	2006
	24AR	Blood culture	1	HIV+	2007
	25AR	Bone biopsy	-	-	2013
	26AR	Abscess at thumb	1	-	2015

Note: (-) indicates that related information was not available.

Abbreviations: HIV, human immunodeficiency virus; TB, tuberculosis.

Table 2 Minimum Inhibitory Concentration of the Antibiotics Tested and Stability Over Time

	Incubation Time (Days)
	7	14	21	28	35
Amikacin	0.5	0.5	0.5	0.5	0.5
Ethambutol	0.625	0.625	0.625	1.25	5
Isoniazid	0.05	0.05	0.05	0.05	0.05
Ofloxacin	0.25	0.25	0.25	0.25	0.25
Rifampicin	0.25	0.25	0.25	0.25	0.25
Streptomycin	0.25	0.25	0.25	0.25	0.25

Note: MIC values are expressed in mg/L.

Table 3 Breakpoints for Broth Micro-Dilution Method for Mycobacterium tuberculosis Complex Published by the CLSI

	Breakpoints (mg/L)
	Susceptible	Inconclusive	Resistant
Ethambutol	≤2	4	≥8
Isoniazid	≤0.12	–	≥0.25
Rifampicin	≤1	–	≥2

Table 4 Drug Susceptibility Testing of BCG Vaccines to First and Second-Line Antituberculosis Drugs

Vaccine	AMK	EMB	INH	OFX	RIF	STR
BCG Moreau	0.5	1.25	0.05	0.5	0.25	0.25
BCG Sofia	0.5	0.625	0.05	0.25	0.25	0.25
BCG Pasteur	0.5	2.5	0.1	0.5	0.25	0.25

Note: MIC values are expressed in mg/L.

Abbreviations: AMK, amikacin; EMB, ethambutol; INH, isoniazid; OFX, ofloxacin; RIF, rifampicin; STR, streptomycin.

Figure 1 Minimum inhibitory concentration of BCG clinical strains for all the drugs tested.

Notes: Pointed lines indicate the breakpoint for resistance according to CLSI. For streptomycin, amikacin and ofloxacin the breakpoints for this method have not yet been determined. Abbreviations: MIC, Minimum inhibitory concentration.

Table 5 Minimum Inhibitory Concentrations of BCG Clinical Strains Against the Antibiotics Tested

BCG Moreau-Related Clinical Strains
	Range	MIC50	MIC90	Mode
Amikacin	0.5–4	0.5	0.5	0.5
Ethambutol	1.25–5	2.5	2.5	2.5
Isoniazid	0.05–1.6	0.1	1.6	0.1–1.6
Ofloxacin	0.25–0.5	0.5	0.5	0.5
Rifampicin	0.25–8	0.25	8	0.25
Streptomycin	0.25	0.25	0.25	0.25
BCG Sofia-Related Clinical Strains
	Range	MIC50	MIC90	Mode
Amikacin	0.5	0.5	0.5	0.5
Ethambutol	0.3125–10	2.5	5	2.5–5
Isoniazid	0.05	0.05	0.05	0.05
Ofloxacin	0.25–0.5	0.25	0.5	0.25–0.5
Rifampicin	0.25	0.25	0.25	0.25
Streptomycin	0.25	0.25	0.25	0.25
BCG Pasteur-Related Clinical Strains
	Range	MIC50	MIC90	Mode
Amikacin	0.5	0.5	0.5	0.5
Ethambutol	0.3125–5	2.5	5	5
Isoniazid	0.05–1.6	0.05	0.05	0.05
Ofloxacin	0.25	0.25	0.25	0.25
Rifampicin	0.25–8	0.25	8	0.25
Streptomycin	0.25	0.25	0.25	0.25

Note: MIC values are expressed in mg/L.

Abbreviation: MIC₅₀ and MIC₉₀, minimum inhibitory concentration at which 50% and 90% of the strains are inhibited.