新生儿百日咳致病株抗菌药物敏感性和抗原基因型研究
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摘要
目的探讨新生儿百日咳抗菌药物的可选方案,阐明致病株的抗原基因型。方法以2013年5月至2018年7月分离到的32株新生儿百日咳鲍特菌为研究对象。采用E-test法检测红霉素、磺胺甲基异噁唑-甲氧苄啶(SMZ)、氨苄西林等共18种抗菌药物的最低抑菌浓度(MIC);扩增分离株23S rRNA基因并测序,检测红霉素耐药基因的突变位点,分析菌株抗原相关的7个基因型(ptxA、ptxC、ptxP、prn、fim2、fim3和tcfA)。结果 25株(25/32,78%)百日咳鲍特菌对红霉素、阿奇霉素、克拉霉素、克林霉素的MIC值均>256 mg/L,且23S rRNA基因均有红霉素耐药A2047G突变。所有菌株对SMZ的MIC值均≤0.064 mg/L。氨苄西林、阿莫西林、阿莫西林‐克拉维酸和头孢曲松的MIC值波动在0.032～1 mg/L。大环内酯类耐药菌株的抗原基因型均为ptxA1/ptxC1/ptxP1/prn1/fim2-1/fim3-1/tcfA2。结论新生儿百日咳鲍特菌对大环内酯类抗菌药物耐药常见,体外试验支持超说明书使用磺胺类抗菌药物是治疗大环内酯类耐药的新生儿百日咳的可靠方案。耐药菌流行更加强调了免疫预防的重要性。
Objective To investigate the alternative antimicrobial drugs for the treatment of neonatal pertussis and the antigen genotypes of Bordetella pertussis(B. pertussis) strains. Methods A total of 32 B. pertussis strains isolated from neonates between May 2013 and July 2018 were used in this study. E-test stripes were used to measure the minimal inhibitory concentration(MIC) of 18 antimicrobial drugs including erythromycin, sulfamethoxazole-trimethoprim(SMZ) and ampicillin. The 23 S rRNA gene of isolated strains was amplified and sequenced to identify the mutation site of erythromycin resistance gene, and the seven antigen genotypes of B. pertussis strains(ptxA, ptxC, ptxP, prn,fim2, fim3 and tcfA2) were analyzed. Results Of the 32 B. pertussis strains, 25(78%) were resistant to erythromycin,azithromycin, clarithromycin and clindamycin, with an MIC of >256 mg/L, and A2047 G mutation was observed in the23 S rRNA gene. All strains had an MIC of ≤0.064 mg/L for SMZ. The MIC of ampicillin, amoxicillin, amoxicillinclavulanic acid and ceftriaxone ranged from 0.032 to 1 mg/L. The strains resistant to macrolide antibiotics had an antigen genotype of ptxA1/ptxC1/ptxP1/prn1/fim2-1/fim3-1/tcfA2. Conclusions B. pertussis strains from neonates are often resistant to macrolides, and the in vitro test shows that off-label use of sulfonamides is a reliable regimen for the treatment of neonates with macrolide-resistant pertussis. The prevalence of drug-resistant strains further emphasizes the importance of immunoprophylaxis.
Objective To investigate the alternative antimicrobial drugs for the treatment of neonatal pertussis and the antigen genotypes of Bordetella pertussis(B. pertussis) strains. Methods A total of 32 B. pertussis strains isolated from neonates between May 2013 and July 2018 were used in this study. E-test stripes were used to measure the minimal inhibitory concentration(MIC) of 18 antimicrobial drugs including erythromycin, sulfamethoxazole-trimethoprim(SMZ) and ampicillin. The 23 S rRNA gene of isolated strains was amplified and sequenced to identify the mutation site of erythromycin resistance gene, and the seven antigen genotypes of B. pertussis strains(ptxA, ptxC, ptxP, prn,fim2, fim3 and tcfA2) were analyzed. Results Of the 32 B. pertussis strains, 25(78%) were resistant to erythromycin,azithromycin, clarithromycin and clindamycin, with an MIC of >256 mg/L, and A2047 G mutation was observed in the23 S rRNA gene. All strains had an MIC of ≤0.064 mg/L for SMZ. The MIC of ampicillin, amoxicillin, amoxicillinclavulanic acid and ceftriaxone ranged from 0.032 to 1 mg/L. The strains resistant to macrolide antibiotics had an antigen genotype of ptxA1/ptxC1/ptxP1/prn1/fim2-1/fim3-1/tcfA2. Conclusions B. pertussis strains from neonates are often resistant to macrolides, and the in vitro test shows that off-label use of sulfonamides is a reliable regimen for the treatment of neonates with macrolide-resistant pertussis. The prevalence of drug-resistant strains further emphasizes the importance of immunoprophylaxis.
引文
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[18]Wang LM,Qiao XL,Ai L,et al.Isolation of antimicrobial resistant bacteria in upper respiratory tract infections of patients[J].3 Biotech,2016,6(2):166.
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[20]Clarke M,McIntyre PB,Blyth CC,et al.The relationship between Bordetella pertussis genotype and clinical severity in Australian children with pertussis[J].J Infect,2016,72(2):171-178.
[21]van Gent M,Heuvelman CJ,van der Heide HG,et al.Analysis of Bordetella pertussis clinical isolates circulating in European countries during the period 1998-2012[J].Eur J Clin Microbiol Infect Dis,2015,34(4):821-830.
[2]World Health Organization.Pertussis vaccines:WHO position paper-August 2015[J].Wkly Epidemiol Rec,2015,90(35):433-460.
[3]Warfel JM,Zimmerman LI,Merkel TJ.Acellular pertussis vaccines protect against disease but fail to prevent infection and transmission in a nonhuman primate model[J].Proc Natl Acad Sci USA,2014,111(2):787-792.
[4]Kim SH,Lee J,Sung HY,et al.Recent trends of antigenic variation in Bordetella pertussis isolates in Korea[J].J Korean Med Sci,2014,29(3):328-333.
[5]Mosiej E,Zawadka M,Krysztopa-Grzybowska K,et al.Sequence variation in virulence-related genes of Bordetella pertussis isolates from Poland in the period 1959-2013[J].Eur JClin Microbiol Infect Dis,2015,34(1):147-152.
[6]Advani A,Gustafsson L,Ahrén C,et al.Appearance of Fim3and ptxP3-Bordetella pertussis strains,in two regions of Sweden with different vaccination programs[J].Vaccine,2011,29(18):3438-3442.
[7]Wiley KE,Zuo Y,Macartney KK,et al.Sources of pertussis infection in young infants:a review of key evidence informing targeting of the cocoon strategy[J].Vaccine,2012,1(4):618-625.
[8]Centers for Disease Control and Prevention.Pertussis surveillance reports[DB/OL].(2014-01-21)[2018-12-27].https://www.cdc.gov/pertussis/surv-reporting.html.
[9]郭丹,李耿.新生儿百日咳[J].儿科学大查房,2015,4(6):265-270.
[10]Abu Raya B,Srugo I,Kessel A,et al.The effect of timing of maternal tetanus,diphtheria,and acellular pertussis(Tdap)immunization during pregnancy on newborn pertussis antibody levels-a prospective study[J].Vaccine,2014,32(44):5787-5793.
[11]Meng QH,Luo J,Yang F,et al.A general lack of IgG against pertussis toxin in Chinese pregnant women and newborns[J].Pediatr Infect Dis J,2018,37(9):934-938.
[12]Tiwari T,Murphy TV,Moran J,et al.Recommended antimicrobial agents for the treatment and postexposure prophylaxis of pertussis:2005 CDC Guidelines[J].MMWRRecomm Rep,2005,54(RR-14):1-16.
[13]Hoppe JE,Haug A.Treatment and prevention of pertussis by antimicrobial agents(Part II)[J].Infection,1988,16(3):148-152.
[14]Islur J,Anglin CS,Middleton PJ.The whooping cough syndrome:a continuing pediatric problem[J].Clin Pediatr(Phila),1975,14(2):171-176.
[15]Trollfors B.Effect of erythromycin and amoxycillin on Bordetella pertussis in the nasopharynx[J].Infection,1978,6(5):228-230.
[16]Simon C,Bontemps M,Wiese K,et al.Ampicillin therapy of whooping cough[J].Dtsch Med Wochenschr,1969,94(47):2435-2437.
[17]Smith BR,LeFrock JL.Bronchial tree penetration of antibiotics[J].Chest,1983,83(6):904-908.
[18]Wang LM,Qiao XL,Ai L,et al.Isolation of antimicrobial resistant bacteria in upper respiratory tract infections of patients[J].3 Biotech,2016,6(2):166.
[19]Ohtsuka M,Kikuchi K,Shimizu K,et al.Emergence of quinolone-resistant Bordetella pertussis in Japan[J].Antimicrob Agents Chemother,2009,53(7):3147-3149.
[20]Clarke M,McIntyre PB,Blyth CC,et al.The relationship between Bordetella pertussis genotype and clinical severity in Australian children with pertussis[J].J Infect,2016,72(2):171-178.
[21]van Gent M,Heuvelman CJ,van der Heide HG,et al.Analysis of Bordetella pertussis clinical isolates circulating in European countries during the period 1998-2012[J].Eur J Clin Microbiol Infect Dis,2015,34(4):821-830.