肺炎链球菌毒力特征及其PBP基因突变和β-内酰胺类耐药情况

doi:10.3969/j.issn.1673-8640.2025.05.006

摘要/Abstract

摘要：

目的了解肺炎链球菌毒力特征及其青霉素结合蛋白（PBP）基因突变和β-内酰胺类耐药情况。方法收集2022年1—12月唐山市妇幼保健院25株肺炎链球菌临床分离株。采用全自动微生物鉴定药敏仪进行菌株鉴定。采用微量肉汤稀释法检测菌株青霉素、阿莫西林、头孢呋辛、头孢曲松钠的最低抑菌浓度（MIC）。采用聚合酶链反应（PCR）检测肺炎链球菌毒力基因（Ply、LytA、NanA、PsaA、PspA）。分析PBP2b基因突变与β-内酰胺类耐药的相关性。结果 25株肺炎链球菌中，有21株（84.00%）分离自痰液样本。有10株（40.00%）对β-内酰胺类抗菌药物敏感；有15株（60.00%）对β-内酰胺类抗菌药物耐药，其中3株MIC值为4~32 μg·mL^-1。25株肺炎链球菌PBP2b阳性检出率为100.00%。低MIC值（1~2 μg·mL^-1）耐药菌株有5株，其丝氨酸-X-Y赖氨酸发生改变；中等MIC值（1~4 μg·mL^-1）耐药菌株有7株，丝氨酸-X-Y赖氨酸、丝氨酸-X-Y天冬酰胺和赖氨酸-苏氨酸/丝氨酸-甘氨酸发生改变；高MIC值（4~32 μg·mL^-1）耐药菌株有3株，PBP2b编码序列保守区SSN附近苏氨酸445/451—丙氨酸/丝氨酸、丝氨酸-甘氨酸发生改变；nanA、LytA和Ply基因阳性检出率均>80%，nanA+Ply+LytA模式阳性率最高（56.00%）。结论肺炎链球菌临床分离株PBP基因编码序列保守区SSN附近苏氨酸445/451—丙氨酸/丝氨酸、丝氨酸-甘氨酸与β-内酰胺类耐药密切相关，且毒力基因nanA、LytA和Ply分布较广。

关键词: 肺炎链球菌, 青霉素结合蛋白, 基因突变, β-内酰胺类抗菌药物, 毒力基因, 耐药

Abstract:

Objective To analyze the virulence characteristics of Streptococcus pneumoniae，penicillin-binding protein （PBP） gene mutation and β-lactam resistance. Methods Totally，25 clinical isolates of Streptococcus pneumoniae were collected from Tangshan Maternal and Child Health Hospital from January to December 2022. The isolates were identified by automatic microbial identification drug susceptibility instrument. The minimum inhibitory concentrations （MIC） of penicillin，amoxicillin，cefuroxime and ceftriaxone sodium were determined by microbroth dilution method. The virulence genes （Ply，LytA，NanA，PsaA and PspA）of Streptococcus pneumoniae were determined by polymerase chain reaction （PCR）. The correlation between PBP2b gene mutation and β-lactam resistance was analyzed. Results Among the 25 isolates of Streptococcus pneumoniae，21 （84.00%） isolates were isolated from sputum samples. Totally，10（40.00%）isolates were sensitive to β-lactam antibiotics. There were 15（60.00%）isolates resistant to β-lactam antibiotics，and the MIC of 3 isolates were 4-32 μg·mL^-1. The PBP2b positive rate of 25 isolates of Streptococcus pneumoniae was 100.00%. There were 5 resistant isolates with low MIC（1-2 μg·mL^-1），and serine-X-Y lysine was changed. There were 7 isolates with medium MIC（1-4 μg·mL^-1），and serine-X-Y lysine，serine-X-Y asparagine and lysine-threonine/serine-glycine were changed. There were 3 isolates with high MIC（4-32 μg·mL^-1），and there were changes of threonine 445/451-alanine/serine and lysune-threonine/serine-glycine in the conserved region of PBP2b coding sequence near SSN. The positive rates of nanA，LytA and Ply were 80.00%，and the positive rate of nanA+Ply+LytA model was the highest（56.00%）. Conclusions Threonine 445/451-alanine/serine and lysune-threonine/serine-glycine are related to β-lactam resistance in the conserved region of PBP2b coding sequence，and the virulence genes（nanA，LytA and Ply）are widely distributed.

Key words: Streptococcus pneumoniae, Penicillin-binding protein, Gene mutation, β-lactam antibiotic, Virulence gene, Drug resistance

中图分类号:

R378.1

姚玲, 高春燕, 刘明雷. 肺炎链球菌毒力特征及其PBP基因突变和β-内酰胺类耐药情况[J]. 检验医学, 2025, 40(5): 450-454.

YAO Ling, GAO Chunyan, LIU Minglei. Virulence characteristics of Streptococcus pneumoniae，PBP gene mutation and β-lactam resistance[J]. Laboratory Medicine, 2025, 40(5): 450-454.

图/表 2

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菌株编号	阿莫西林	青霉素	头孢呋辛	头孢曲松钠	判定结果
1、3、14号	0.14	0.14	0.05	0.14	敏感
9、17、19号	0.20	0.20	0.10	0.10	敏感
5、7、12、21号	0.80	0.80	0.40	0.40	敏感
11、23号	1.00	1.00	1.00	1.00	耐药
2、15号	1.00	2.00	1.00	1.00	耐药
6、13号	4.00	2.00	1.00	1.00	耐药
4号	2.00	4.00	1.00	2.00	耐药
8号	2.00	4.00	2.00	4.00	耐药
10号	2.00	2.00	1.00	2.00	耐药
16号	2.00	4.00	1.00	1.00	耐药
18、22号	2.00	4.00	2.00	4.00	耐药
20号	4.00	8.00	32.00	4.00	耐药
24、25号	16.00	8.00	8.00	4.00	耐药

菌株编号	阿莫西林	青霉素	头孢呋辛	头孢曲松钠	判定结果
1、3、14号	0.14	0.14	0.05	0.14	敏感
9、17、19号	0.20	0.20	0.10	0.10	敏感
5、7、12、21号	0.80	0.80	0.40	0.40	敏感
11、23号	1.00	1.00	1.00	1.00	耐药
2、15号	1.00	2.00	1.00	1.00	耐药
6、13号	4.00	2.00	1.00	1.00	耐药
4号	2.00	4.00	1.00	2.00	耐药
8号	2.00	4.00	2.00	4.00	耐药
10号	2.00	2.00	1.00	2.00	耐药
16号	2.00	4.00	1.00	1.00	耐药
18、22号	2.00	4.00	2.00	4.00	耐药
20号	4.00	8.00	32.00	4.00	耐药
24、25号	16.00	8.00	8.00	4.00	耐药

毒力基因组合模式	菌株数/株	百分比/%
nanA+LytA	2	8.00
nanA+Ply	1	4.00
nanA+Ply+LytA	14	56.00
nanA+Ply+LytA+PspA	2	8.00
nanA+Ply+LytA+PsaA	5	20.00
nanA+Ply+LytA+PsaA+PspA	1	4.00

毒力基因组合模式	菌株数/株	百分比/%
nanA+LytA	2	8.00
nanA+Ply	1	4.00
nanA+Ply+LytA	14	56.00
nanA+Ply+LytA+PspA	2	8.00
nanA+Ply+LytA+PsaA	5	20.00
nanA+Ply+LytA+PsaA+PspA	1	4.00