多重聚合酶链反应在肺炎链球菌血清学分型中的应用

doi:10.3969/j.issn.1673-8640.2014.11.003

摘要/Abstract

摘要： 肺炎链球菌是儿科感染中常见的病原菌之一。基于荚膜多糖抗原的特点,传统荚膜肿胀实验可将肺炎链球菌分为46个血清群,90多种血清型。近年来随着分子生物学技术的发展,多重聚合酶链反应(MP-PCR)技术不断成熟并逐步代替常规的荚膜肿胀实验,用于血清学分型。该技术具有简便快速、成本相对较低等优点,能有效的监测临床上肺炎链球菌流行血清型。

关键词: 肺炎链球菌, 多重聚合酶链反应, 血清型

Abstract: Streptococcus pneumoniae is one of common pathogens causing pediatric infections. Owing to the characteristic of capsular polysaccharide, Streptococcus pneumoniae can be differentiated into 40 serogroups and more than 90 serotypes by conventional capsular quellung reaction. Recently, with the development of molecular biological techniques, multiplex polymerase chain reaction (MP-PCR) gradually replaces the conventional capsular quellung reaction, which is used to detect the serotypes of Streptococcus pneumoniae. Furthermore, MP-PCR has the advantages of easy and rapid operation and low costing, which could provide the basis for monitoring the prevalent serotypes of Streptococcus pneumoniae in clinic.

Key words: Multiplex polymerase chain reaction, Serotype

中图分类号:

R446.1

潘芬，张泓. 多重聚合酶链反应在肺炎链球菌血清学分型中的应用[J]. 检验医学, 2014, 29(11): 1089-1093.

PAN Fen, ZHANG Hong. Application of multiplex polymerase chain reaction for detecting the serotypes of Streptococcus pneumonia[J]. , 2014, 29(11): 1089-1093.

参考文献

[1] Mavroidi A, Aanensen DM, Godoy D, et al. Genetic relatedness of the Streptococcus pneumoniae capsular biosynthetic loci [J]. J Bacteriol, 2007, 189 (21): 7841-7855.
[2] Sørensen UB. Typing of pneumococci by using 12 pooled antisera [J]. J Clin Microbiol, 1993, 31 (8): 2097-2100.
[3] Pai R, Gertz RE, Beall B. Sequential multiplex PCR approach for determining capsular serotypes of Streptococcus pneumoniae isolates [J]. J Clin Microbiol, 2006, 44 (1): 124-131.
[4] Elberse KE, van de Pol I, Witteveen S, et al. Population structure of invasive Streptococcus pneumoniae in the Netherlands in the pre-vaccination era assessed by MLVA and capsular sequence typing [J]. PLoS One, 2011, 6 (5): e20390.
[5] Wang Q, Wang M, Kong F,el al. Development of DNA microarray to identify the Streptococcus pneumoniae serotypes contained in the 23-valent pneumococcal polysaccharide vaccine and closely related serotypes [J]. J Microbiol Methods, 2007, 68 (1): 128-136.
[6] Bentley SD, Aanensen DM, Mavroidi A, et al. Genetic analysis of the capsular biosynthetic locus from all 90 pneumococcal serotypes [J]. PLoS Genet, 2006, 2 (3): e31.
[7] Cartee RT, Forsee WT, Jensen JW, et al. Expression of the Streptococcus pneumoniae type 3 synthase in Escherichia coli . Assembly of type 3 polysaccharide on a lipid primer [J]. J Biol Chem, 2001, 276 (52): 48831-48839.
[8] Dillard JP, Vandersea MW, Yother J. Charac-terization of the cassette containing genes for type 3 capsular polysaceharide biosynthesis in Streptococcus pneumoniae [J]. J Exp Med, 1995, 18l (3): 973-983.
[9] Kolkman MA, van der Zeijst BA, Nuijten PJ. Diversity of capsular polysaccharide synthesis gene clusters in Streptococcus pneumoniae [J]. J Biochem, 1998, 123 (5): 937-945.
[10] García E, Llull D, Muñoz R, et al. Current trends in capsular polysaccharide biosynthesis of Streptococcus pneumoniae [J]. Res Microbiol, 2000, 151 (6): 429-435.
[11] Lawrence ER, Griffiths DB, Martin SA, et al. Evaluation of semiautomated multiplex PCR assay for determination of Streptococcus pneumoniae serotypes and serogroups [J]. J Clin Microbiol, 2003, 41(2): 601-607.
[12] Brito DA, Ramirez M, de Lencastre H. Serotyping Streptococcus pneumoniae by multiplex PCR [J]. J Clin Microbiol, 2003, 41(6): 2378-2384.
[13] Ahn JG, Choi SY, Kim DS, et al. Enhanced detection and serotyping of Streptococcus pneumoniae using multiplex polymerase chain reaction [J]. Korean J Pediatr, 2012, 55(11): 424-429.
[14] Iraurgui P, Torres MJ, Gandia A, et al. Modified sequential multiplex PCR for determining capsular serotypes of invasive pneumococci recovered from Seville [J]. Clin Microbiol Infect, 2010, 16(9): 1504-1507.
[15] Siira L, Kaijalainen T, Lambertsen L, et al. From quellung to multiplex PCR, and back when needed, in pneumococcal serotyping [J]. J Clin Microbiol, 2012, 50(8): 2727-2731.
[16] Pimenta FC, Roundtree A, Soysal A, et al. Sequential triplex real-time PCR assay for detecting 21 pneumococcal capsular serotypes that account for a high global disease burden [J]. J Clin Microbiol, 2013, 51(2): 647-652.
[17] Zhou F, Kong F, Tong Z, et al. Identification of less-common Streptococcus pneumoniae serotypes by a multiplex PCR-based reverse line blot hybridization assay[J]. J Clin Microbiol, 2007, 45(10): 3411-3415.
[18] Pimenta FC, Gertz RE Jr, Roundtree A, et al. Rarely occurring 19A-like cps locus from a serotype 19F pneumococcal isolate indicates continued need of serology-based quality control for PCR-based serotype determinations [J]. J Clin Microbiol, 2009, 47(7): 2353-2354.
[19] Vickers I, O'Flanaqan D, Cafferkey M, et al. Multiplex PCR to determine Streptococcus pneumoniae serotypes causing otitis media in the Republic of Ireland with further characterisation of antimicrobial susceptibilities and genotypes [J]. Eur J Clin Microbiol Infect Dis, 2011, 30(3): 447-453.
[20] Ercibengoa M, Arostegi N, Marimón JM, et al. Dynamics of pneumococcal nasopharyngeal carriage in healthy children attending a day care center in northern Spain. Influence of detection techniques on the results [J]. BMC Infect Dis, 2012, 12:69.
[21] Castañeda E, Agudelo CI, Regueira M, et al. Laboratory-based surveillance of Streptococcus pneumoniae invasive disease in children in 10 Latin American countries: a SIREVA Ⅱ project, 2000-2005 [J]. Pediatr Infect Dis J, 2009, 28(9): e265-e270.
[22] Morais L, Carvalho Mda G, Roca A,et al. Sequential multiplex PCR for identifying pneumococcal capsular serotypes from South-Saharan African clinical isolates [J]. J Med Microbiol, 2007, 56(Pt 9): 1181-1184.
[23] Zhang YJ, Chen YS, Wang ZW, et al. Serological and molecular capsular typing, antibiotic susceptibility and multilocus sequence typing of Streptococcus pneumoniae isolates from invasive and non-invasive infections [J]. Chin Med J (Engl), 2013, 126(12): 2296-2303.
[24] Weinberger DM, Malley R, Lipsitch M. Serotype replacement in disease after pneumococcal vaccination [J]. Lancet, 2011, 378(9807): 1962-1973.
[25] Hanna-Wakim R, Chehab H, Mahfouz I, et al. Epidemiologic characteristics, serotypes, and antimicrobial susceptibilities of invasive Streptococcus pneumoniae isolates in a nationwide surveillance study in Lebanon [J]. Vaccine, 2012, 30 (Supple 6): G11-G17.
[26] Dos Santos SR, Passadore LF, Takaqi EH, et al. Serotype distribution of Streptococcus pneumoniae isolated from patients with invasive pneumococcal disease in Brazil before and after ten-pneumococcal conjugate vaccine implementation [J]. Vaccine, 2013, 31(51): 6150-6154.
[27] Ho PL, Chiu SS, Ang I, et al. Serotypes and antimicrobial susceptibilities of invasive Streptococcus pneumoniae before and after introduction of 7-valent pneumococcal conjugate vaccine, Hong Kong, 1995-2009 [J]. Vaccine, 2011, 29(17): 3270-3275.
[28] Ho PL, Chiu SS, Chan MY, et al. Changes in nasopharyngeal carriage and serotype distribution of antibiotic-resistant Streptococcus pneumoniae before and after the introduction of 7-valent pneumococcal conjugate vaccine in Hong Kong [J].Diagn Microbiol Infect Dis, 2011, 71(4): 327-334.
[29] Saha SK, Darmstadt GL, Baqui AH, et al. Identification of serotype in culture negative pneumococcal meningitis using sequential multiplex PCR: implication for surveillance and vaccine design [J]. PLoS One, 2008, 3(10): e3576.
[30] O'Brien KL, Millar EV, Zell ER, et al. Effect of pneumococcal conjugate vaccine on nasopharyngeal colonization among immunized and unimmunized children in a community- randomized trial [J]. J Infect Dis, 2007, 196(8): 1211-1120.
[31] Jourdain S, Drèze PA, Vandeven J, et al. Sequential multiplex PCR assay for determining capsular serotypes of colonizing S.pneumoniae [J]. BMC Infect Dis, 2011, 11:100.