Molecular Characterization of Klebsiella pneumoniae Isolated from Sputum in a Tertiary Hospital in Xinxiang, China

Figures & data

Table 1 PCR Primer Sequence Information of Antimicrobial Resistance Genes

Gene	Primer Sequence	Product Size (bp)	Ta Opt (°C)	References
blaCTX-M	P1: TCGGGAGGCAGACTGGGTGT	688	57.7
	P2: CCTTAGGTTGAGGCTGGGTGA
blaTEM	P1: TCGCCGCATACACTATTCTCAG	445	55.1	[42]
	P2: ACGCTCACCGGCTCCAGATTTAT
blaSHV	P1: ATGCGTTATATTCGCCTGTG	753	58.4	[42]
	P2: TGCTTTGTTATTCGGGCCAA
blaKPC	P1: ATGTCACTGTATCGCCGTC	882	57.0
	P2: TTACTGCCCGTTGACGCC
blaIMP	P1: CTTGATGAAGGCGTTTATGT	496	50.9
	P2: GCCAAGCTTCTATATTTGCGT
blaNDM	P1: CCAGCTCGCACCGAATG	564	58.8
	P2: AACGCCGCACCAAACG
blaOXA-48	P1: ACATAAATCACAGGGCGTAG	500	51.4
	P2: TATAGTCACCATTGGCTTCG
qnrA	P1: CAAGAGGATTTCTCACGCCAGGAT	521	58.9
	P2: TCGGCAAAGGTCAGGTCACAGC
qnrB	P1: GCACTGAATTTATCGGCTGTC	500	53.4
	P2: CAACGATGCCTGGTAGTTGT
oqxA	P1: TCCATACCAACCTCGTCTCCC	529	59.8
	P2: AGCGTGGCTTTGAACTCTGC
oqxB	P1: TGGTGGAGAACGTCGAGCGTAA	648	60.4
	P2: TCGGCGTGTTGGTGAACTGC
aac	P1: AGTACAGCATCGTGACCAACA	545	55.8	[43]
	P2: CTCGAATGCCTGGCGTGTTT

Abbreviations: PS, primer sequences without reference were obtained by primer 5.0 design (same below); bp, base pair; Ta opt, the optimal annealing temperature.

Table 2 PCR Primers Sequence Information of Virulence Genes

Gene	Primer Sequence	Product Size (bp)	Ta Opt (°C)	References
wzi	P1: GTGCCGCGAGCGCTTTCTATC TTG GTATTCC	580	55	[16]
	P2: GAGAGCCACTGGTTCCAGAA[CorT]TT[CorG]ACCGC
magA	P1: GGTGCTCTTTACATCATTGC	1282	58	[44]
	P2: GCAATGGCCATTTGCGTTAG
rmpA	P1: ACTGGGCTACCTCTGCTTCA	516	58	[36]
	P2: CTTGCATGAGCCATCTTTCA
iucA	P1: CCCGCTTCCCTACTTT	575	54.8
	P2: ATTCGCTTCGCTGTCC
iutA	P1: GGCTGGACATCATGGGAACTGG	300	55	[36]
	P2: CGTCGGGAACGGGTAGAATCG
wabG	P1: ACCATCGGCCATTTGATAGA	683	49	[36]
	P2: CGGACTGGCAGATCCATATC
fimH	P1: TGCTGCTGGGCTGGTCGATG	688	49	[36]
	P2: GGGAGGGTGACGGTGACATC
markD	P1: TTCTGCACAGCGGTCCC	480	49	[36]
	P2: GATACCCGGCGTTTTCGTTAC

Abbreviations: bp, base pair; Ta opt, the optimal annealing temperature.

Figure 1 The resistant rates of 20 K. pneumoniae isolated from sputum against 13 antibiotics.

Abbreviations: CAZ, Ceftazidime; CTX, Cefotaxime; CFX, Cefoxitin; PIP, Piperacillin; AZT, Aztreonam; PIT, Piperacillin-tazobactam; CHL, Chloramphenicol; IPN, Imipenem; DOX, Doxycycline; KAN, Kanamycin; GAT, Gatifloxacin; LVF, Levofloxacin; CIP, Ciprofioxacin; MDR, multidrug-resistant.

Table 3 Distribution of Antibiotic Resistance Genes and ESBL Phenotype in 20 Strains of K. pneumoniae

Resistance Genes	Distribution
	Number of Strains (n=20)	Resistant Rates (%)
ESBL phenotype	11	55.0
blaSHV	12	60.0
blaCTX-M	7	35.0
blaTEM	8	40.0
blaKPC	3	15.0
blaNDM	0	0.0
blaOXA-48	0	0.0
blaIMP	0	0.0
qnrA	0	0.
qnrB	5	25.0
oqxA	18	90.0
oqxB	19	95.0
aac	8	40.0
blaKPC+blaSHV+oqxAB	2	10.0
blaCTX-M+blaTEM+aac+oqxAB	6	30.0

Table 4 Distribution of Virulence Genes of K. pneumoniae Isolated from Sputum

Distribution	Virulence Phenotypes		Virulence Genes
	BF	HMV	wzi	rmpA	magA	wabG	fimH	mrkD	iucA	iutA
Number of positive strains	20	5	20	5	2	20	18	17	7	6
Positive rates (%)	100	25	100.0	25.0	10.0	100.0	90.0	85.0	35.0	30.0

Abbreviations: BF, biofilm; HMV, hypermucoviscous phenotype.

Figure 2 Phylogenetic tree of 20 K. pneumoniae isolated from sputum base on wzi gene sequence. Phylogenetic trees were constructed using MEGA 7 based on the neighbor-joining method (500 bootstrap replicates) and Jukes-Cantor distance. Each sequence corresponds indicated a distinct wzi allele. The corresponding capsular types followed the allele number.

Figure 3 Pearson’s correlation analysis between phenotypes and genotypes of 20 K. pneumoniae isolated from sputum. Red is positive correlations and blue is negative correlations between different parameters.

Abbreviations: CR, Carbapenem-like antibiotic resistance phenotype; QR, Quinolone antibiotic resistance phenotype; AR, Aminoglycoside antibiotic resistance phenotype; SBF, Strong biofilm.

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