Resistance mechanisms and molecular epidemiology of Pseudomonas aeruginosa from patients with bronchiectasis in Shanghai

doi:10.3969/j.issn.1673-8640.2023.11.003

Abstract

Abstract:

Objective To investigate the drug resistance and molecular epidemiological characteristics of Pseudomonas aeruginosa（PA） isolated from patients with bronchiectasis in Shanghai. Methods Totally，73 isolates of PA from sputum samples of patients with bronchiectasis in 6 hospitals in Shanghai were collected. The durg susceptibility was analyzed by microbroth dilution method. The resistance mechanisms were assessed by next generation sequencing（NGS），and the molecular epidemiological characteristics were determined by multilocus sequence typing（MLST）. Results A total of 73 isolates of PA were collected，which included 43 mucous PA（58.9%）. The highest resistance rates were observed for levofloxacin（41.1%），ciprofloxacin（34.2%） and imipenem（12.3%），and the resistance rates to other antibiotics were all <6%. The most frequent mutation sites of PA which were resistant to fluoroquinolones were concentrated on the 83rd and 87th sites of GyrA，the 466th site of GyrB，the 646th and the 262nd sites of ParC and the 533rd site of ParE. All the outer membrane porin D（OprD） of PA which were resistant to imipenem were mutated or missed. Multiple acquired resistance genes were determined，which included beta-lactamases（bla_IMP-45，bla_PAO，bla_OXA-1，bla_OXA-50 and so on），aminoglycosides [aph（3'）-Iib，aac（6'）-Ib3，aac（6'）-Ib-cr，aadA2b]，fosfomycin（fosA），amphetamines（catB7，catB3），sulfonamides（sul1，qacE），quinolones（crpP）. A total of 53 sequence types（ST） were determined. The most common ST was ST277（5.5%，4/73）. Conclusions Most of PA isolated from sputum samples of bronchiectasis patients in Shanghai are mucous types. These isolates are highly susceptible to common used antibiotics except fluoroquinolones. Multiclonality discloses no evidence on spread of specific types of PA in Shanghai.

Key words: Pseudomonas aeruginosa, Mucous type, Resistance, Multilocus sequence typing, Bronchiectasis

CLC Number:

R378.99

CHEN Qing, XU Rong, SUN Jingyong. Resistance mechanisms and molecular epidemiology of Pseudomonas aeruginosa from patients with bronchiectasis in Shanghai[J]. Laboratory Medicine, 2023, 38(11): 1020-1025.

Figures/Tables 4

References 18

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抗菌药物	PA（73株）		黏液型PA（43株）		非黏液型PA（30株）		P值
抗菌药物	耐药	敏感	耐药	敏感	耐药	敏感	P值
左氧氟沙星	41.1	41.1	46.5	30.2	33.3	56.7	0.260
环丙沙星	34.2	47.9	37.2	39.5	30.0	60.0	0.523
亚胺培南	12.3	82.2	7.0	86.0	20.0	76.7	0.192^①
头孢吡肟	5.5	91.8	9.3	86.0	0	100.0	0.232^①
哌拉西林	4.1	94.5	7.0	90.7	0	100.0	0.380^①
头孢他啶	4.1	94.5	7.0	93.0	0	96.7	0.380^①
哌拉西林-他唑巴坦	2.7	91.8	4.7	90.7	0	93.3	0.509^②
美罗培南	2.7	91.8	2.3	90.7	3.3	93.3	1.000^②
妥布霉素	1.4	98.6	2.3	97.7	0	100.0	1.000^②
阿米卡星	0	98.6	0	100.0	0	96.7
多黏菌素B	0	100.0	0	100.0	0	100.0

抗菌药物	PA（73株）		黏液型PA（43株）		非黏液型PA（30株）		P值
抗菌药物	耐药	敏感	耐药	敏感	耐药	敏感	P值
左氧氟沙星	41.1	41.1	46.5	30.2	33.3	56.7	0.260
环丙沙星	34.2	47.9	37.2	39.5	30.0	60.0	0.523
亚胺培南	12.3	82.2	7.0	86.0	20.0	76.7	0.192^①
头孢吡肟	5.5	91.8	9.3	86.0	0	100.0	0.232^①
哌拉西林	4.1	94.5	7.0	90.7	0	100.0	0.380^①
头孢他啶	4.1	94.5	7.0	93.0	0	96.7	0.380^①
哌拉西林-他唑巴坦	2.7	91.8	4.7	90.7	0	93.3	0.509^②
美罗培南	2.7	91.8	2.3	90.7	3.3	93.3	1.000^②
妥布霉素	1.4	98.6	2.3	97.7	0	100.0	1.000^②
阿米卡星	0	98.6	0	100.0	0	96.7
多黏菌素B	0	100.0	0	100.0	0	100.0

蛋白突变位点	突变类型	突变株数/株	突变率/%
GyrA
83	Thr→Ile	7	15.9
83、87	Thr-83→Ala、Asp-87→Asn	1	2.3
87	Asp→Asn	5	11.4
87	Asp→Gly	2	4.5
87	Asp→Tyr	1	2.3
87、805	Asp-87→Asn、Phe-805→Leu	1	2.3
87合并缺失	Asp→Tyr合并缺失	1	2.3
87合并缺失	Asp→Asn合并缺失	1	2.3
679	Asp→Tyr	2	4.5
75	Gly→Ser	1	2.3
231合并缺失	Glu→Lys合并缺失	1	2.3
791合并缺失	Ala→Val合并缺失	1	2.3
82合并插入	Asp→Asn合并插入	1	2.3
	缺失	4	9.1
	插入	1	2.3
	无突变	14	31.8
GyrB
466	Ser→Phe	4	9.1
466	Phe→Tyr	1	2.3
148	His→Asn	2	4.5
453	Leu→Phe	1	2.3
468	Glu→Asp	1	2.3
469	Val→Phe	1	2.3
632	Ala→Thr	1	2.3
721	Met→Ile	1	2.3
749	Pro→Ser	1	2.3
371、466	371Lys→Arg、466Ser→Phe	1	2.3
	无突变	30	68.2
ParC
646	Val→Leu	7	15.9
262	His→Gln	4	9.1
87	Ser→Leu	1	2.3
148	Val→Gly	1	2.3
187	Val→Met	1	2.3
513	Glu→Asp	1	2.3
197、498	Ser-197→Leu、Glu-498→Asp	1	2.3
262、301	His-262→Gln、Arg-301→Cys	1	2.3
284、646	Lys-284→Glu、Val646→Leu	1	2.3
	无突变	26	59.1
ParE
533	Asp→Glu	4	9.1
33	His→Pro	1	2.3
142	Asp→Asn	1	2.3
188	Lys→Thr	1	2.3
346	Ser→Thr	1	2.3
419	Gly→Asn	1	2.3
452	Gly→Ser	1	2.3
492	Ser→Tyr	1	2.3
225、425	Ala-225→Val、Ala-425→Val	1	2.3
472、533	Ala-472→Val、Asp-533→Glu	1	2.3
473、533	Ala-473→Val、Asp-533→Glu	1	2.3
533、599	Asp-533→Glu、Leu-599→Val	1	2.3
473、501、533	Ala-473→Val、Leu-501→Phe、Asp533→Glu	1	2.3
	插入	1	2.3
	无突变	27	61.4

蛋白突变位点	突变类型	突变株数/株	突变率/%
GyrA
83	Thr→Ile	7	15.9
83、87	Thr-83→Ala、Asp-87→Asn	1	2.3
87	Asp→Asn	5	11.4
87	Asp→Gly	2	4.5
87	Asp→Tyr	1	2.3
87、805	Asp-87→Asn、Phe-805→Leu	1	2.3
87合并缺失	Asp→Tyr合并缺失	1	2.3
87合并缺失	Asp→Asn合并缺失	1	2.3
679	Asp→Tyr	2	4.5
75	Gly→Ser	1	2.3
231合并缺失	Glu→Lys合并缺失	1	2.3
791合并缺失	Ala→Val合并缺失	1	2.3
82合并插入	Asp→Asn合并插入	1	2.3
	缺失	4	9.1
	插入	1	2.3
	无突变	14	31.8
GyrB
466	Ser→Phe	4	9.1
466	Phe→Tyr	1	2.3
148	His→Asn	2	4.5
453	Leu→Phe	1	2.3
468	Glu→Asp	1	2.3
469	Val→Phe	1	2.3
632	Ala→Thr	1	2.3
721	Met→Ile	1	2.3
749	Pro→Ser	1	2.3
371、466	371Lys→Arg、466Ser→Phe	1	2.3
	无突变	30	68.2
ParC
646	Val→Leu	7	15.9
262	His→Gln	4	9.1
87	Ser→Leu	1	2.3
148	Val→Gly	1	2.3
187	Val→Met	1	2.3
513	Glu→Asp	1	2.3
197、498	Ser-197→Leu、Glu-498→Asp	1	2.3
262、301	His-262→Gln、Arg-301→Cys	1	2.3
284、646	Lys-284→Glu、Val646→Leu	1	2.3
	无突变	26	59.1
ParE
533	Asp→Glu	4	9.1
33	His→Pro	1	2.3
142	Asp→Asn	1	2.3
188	Lys→Thr	1	2.3
346	Ser→Thr	1	2.3
419	Gly→Asn	1	2.3
452	Gly→Ser	1	2.3
492	Ser→Tyr	1	2.3
225、425	Ala-225→Val、Ala-425→Val	1	2.3
472、533	Ala-472→Val、Asp-533→Glu	1	2.3
473、533	Ala-473→Val、Asp-533→Glu	1	2.3
533、599	Asp-533→Glu、Leu-599→Val	1	2.3
473、501、533	Ala-473→Val、Leu-501→Phe、Asp533→Glu	1	2.3
	插入	1	2.3
	无突变	27	61.4