Pyrosequencing for rapid detection of Tuberculosis resistance in clinical isolates and Sputum samples from re-treatment Pulmonary Tuberculosis patients

Table 1 Primers and thermo-cycling conditions of the pyrosequencing assays

Drug	Gene	Primer	Sequences (5’—3’)	Annealing temperature (°C)	Size (bp)	Target loci
RIF	rpoB	Forward	GTCCGGGAGCGGATGACCACCC	65	204	RRDR
		Reverse*	GCTCACGTGACAGACCGCCG
		Sequencing 1	GCGATCAAGGAGTTC
		Sequencing 2	TCATGGACCAGAACAA
INH	katG	Forward	AGATGGGCTTGGGCTGGA	61	133	315
		Reverse*	TAGCCGTACAGGATCTCGAGGA
		Sequencing	CCGGTAAGGACGCGA
SM	rpsl	Forward*	CAAGGGTCGTCGGGACAAGA	61	299
		Reverse	TCTTGACACCCTGCGTATCC
		Sequencing 1	CGCCGAGTTCGGCTT			43
		Sequencing 2	CACACCAGGCAGGTC			88
EMB	embB	Forward	ACGACGGCTACATCCTGG	61	110	306
		Reverse*	GTTGTAATACCAGCCGAAGGGA
		Sequencing	CGACGGCTACATCCTG
OFL	gyrA	Forward	TTCGATTCCGGCTTCCGCCC	61	192	QRDR
		Reverse*	TGGGTCATTGCCTGGCGAGC
		Sequencing	TACCACCCGCACGGC
AMK	rrs	Forward	TCCTTAAAAGCCGGTCTCAGTTC	61	239	1401
		Reverse*	TCCGGTACGGCTACCTTGTTA
		Sequencing	CTTGTACACACCGCC

ISSN: 1471-2334