PCR diagnostics of Mycobacterium tuberculosis in historic human long bone remains from 18th century burials in Kaiserebersdorf, Austria

Table 1 Primers used for amplifying M. tuberculosis target DNA

Primer	Length of product	Sequence 5' – 3'	Reference
IS1081_F2	135 bp	CTGCTCTCGACGTTCATCGCCG	Insertion sequence IS1081
IS1081_R2		GGCACGGGTGTCGAAATCACG	[2]
IS1081_R3	113 bp	TGGCGGTAGCCGTTGCGC
OxyR_F3	110 bp	CACTGCCTACGCGACCAGACG	oxidative stress regulator OxyR pseudogene
OxyR _R1		TCTGCGGAATCAGTGTCACC	[2]
OxyR _F4	94 bp	AGACGCTCGATGCTGCCCAA
pncA_F2	117 bp	GGCGGACTACCATCACGTCG	Pyrazinamidase gene
pncA _R2		GGAGTACCGCTGACGCAATGCGGT	[2]
pncA _R3	96 bp	GGCCACGACGAGGAATAGTC
D1flanking_F	112 bp	ATCGAAAGGCTAACGGGTGC	flanking sections of TbD1 region
D1flanking _R2		CTGCTCGCCGAGTTGGTCGATA	[2]
D1flanking _R3	96 bp	TCGATACCGTCGATCGTGTCAAAG
D1internal _F3	115 bp	CGAGCTACGACGCTCGCTATTA	TbD1 region
D1internal _R3		GCATATCGTGATCCGTCTCGAT	[2]
D1internal _R4	105 bp	CTCGATCATCAGTAGTTCGGGATTC
TbA	TbA/TbB:	CTCGTCCAGCGCCGCTTCGG	insertion sequence IS6110
TbB	120 bp	CCTGCGAGCGTAGGCGTCGG	[1]
TbC	TbC/TbD:	GCTTCGGACCACCAGCACCT
TbD	95 bp	GCGTCGGTGACAAAGGCCAC

For the primers of [2], F (forward) and R (reverse) denote orientation of the primer and for each gene amplification was performed with the first two primers followed by reamplification with the third primer in combination with one of the original primers.
Target Sequences: IS108 = Insertion sequence IS1081; OxyR = oxidative stress regulator OxyR pseudogene; pncA = Pyrazinamidase gene; D1 = TbD1 region described in [25] that is deleted in modern strains of M. tuberculosis; TbA-TbD: insertion sequence IS6110.

ISSN: 1756-0500