In-vivo loss of carbapenem resistance by extensively drug-resistant Klebsiella pneumoniae during treatment via porin expression modification


Supplementary table (Table

 S1). List 



  




  of primers used in this study Landman, D., George, A., Salvani, J. & Quale, J. Correlation of the expression of acrB and the regulatory genes marA, soxS and ramA with antimicrobial resistance in clinical isolates of Klebsiella pneumoniae endemic to New York City.J Antimicrob Chemother 64,278-283 (2009). 


	ompK36	ompK36RTF	GCGACCAGACCTACATGCGT	Bratu et al.
		ompK36RTR	AGTCGAAAGAGCCCGCGTC	
	Assay and target region phoE	Primer phoEF	Sequence (5'-3') TGATGGGCTTTGTGGCTTC	Reference Bratu et al.
	Sequencing	phoER	CTTGCTGTCATAGTCGCTG	
	ompK35 rpoB	ompK35F rpoBF	CAGACACCAAACTCTCATCA AAGGCGAATCCAGCTTGTTCAGC	This work Bratu et al.
		ompK35R rpoBR	TCAGGTTAACGGGAGAATAA TGACGTTGCATGTTCGCACCCATCA	
		ompK35sqF	GCGTCGATCAGAAAGCT	
	Bratu, S.,	ompK35sqR	GCTTCGGCTTTGTCGCCA	
	ompK36	ompK36F	TTGCAGCACAATGAAATAGC	This work
		ompK36R	CAAGAGTATACCAGCGAGGT	
		ompK36sqF	CGACGTTCTGCCGGAATT	
		ompK36sqR	AGAGTTACGGTAGGTTGC	
	Inter rarA and oxqA	rarA+intF	ACGGACCGTTCACTATGTCG	37
		rarA+intR	GACGAGCAGAAAGGAGAGCA	
	Sequencing and cloning			
	oqxR	oqxRF	TTCCTGACGCCGGTGTTTTA	37
		oqxRR	CTGCGGTGCCAAAAAGAACA	
	pstS	pstSF	CGGCGGTAGATGACAAACGA	This work
		pstSR	CCCCGCCATCTTCGTCAAAA	
		pstSsqF	AAAAATGGGATGACGAAGCG	
		pstSsqR	GGCGCGGCGGACCACGGCGA	
	phoU	phoUF	CGTGACTCACAACATGCAGC	This work
		phoUR	GCCGCGGGGTAAAACATATC	
	phoB	phoBF	CGTCGTGCTTCAAAATACCC	This work
		phoBR	TTCCATGACAGCCGTTCCAG	
	phoR	phoRF	TGGCCATGATCGTATGGTGC	This work
		phoRR	GCAGCGCACGCTCATTAAATA	
		phoRsqF	CGGAACTTTTTCGCCAACGT	
		phoRsqR	CTGCTCCTGCATCATCTCCA	
	RT-PCR			
	acrB	acrBF	CGATAACCTGATGTACATGTCC	Bratu et al.
		acrBR	CCGACAACCATCAGGAAGCT	
	ramA	ramAF	ATCGTCGAGTGGATTGATGA	32
		ramAR	AGATGCCATTTCGAATACCC	
	oqxB	oqxB1806F	GAGCGAGATCGGGATGAATAC	37
		oqxB1890R	CGGCGTGTTGGTGAACTG	
	rarA	Kp02968F	TGGATCGACAACCATCTTGA	23
		Kp02968R	AAGGACTGCTGGGAGTCAAA	
	ompK35	ompK35RTF	TCCCTGCCCTGCTGGTAG	Bratu et al.
		ompK35RTR	CTGGTGTCGCCATTGGTGG	

Supplementary figure (Figure

 S1). Immunodetection of porins and OmpA. After SDS-PAGE separation of denatured samples, the detection was performed with the polyclonal antibodies directed against denatured OmpF (OmpK35 in Klebsiella pneumoniae) porin and also against denatured OmpC (OmpK36 in K. pneumoniae) porin due to cross-recognition, and OmpA. The tested strains, ATCC13883, BJ-STC-a and BJ-STC-b, were grown in Mueller Hinton II (MH2), nutrient broth (NB) and nutrient broth containing sorbitol (NBS) media. A mix of anti-OmpA and anti-OmpF was used on the same nitrocellulose sheet to detect porins and OmpA.
Suzanne Bialek-Davenet, Noémie Mayer, Julia Vergalli, Marion Duprilot, Sylvain Brisse, Jean-Marie Pagès, Marie-Hélène Nicolas-Chanoine
In-vivo loss of carbapenem resistance by extensively drug-resistant Klebsiella pneumoniae during treatment via porin expression modification