肺炎支原体核酸分子诊断技术研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2024-0119

摘要/Abstract

摘要：

肺炎支原体是目前发现的体积最小的细胞生物，也是引发社区获得性肺炎主要的病原体之一。在感染早期，临床表现千差万别，可累及全身各个器官。早期诊断具有一定的挑战性，放射学X线检查和早期临床表现均不具有特异性，易因误诊和漏诊对人们的身体健康带来危害。目前国内外肺炎支原体的检查主要依靠实验室诊断手段。本文从肺炎支原体的复杂发病机制入手，包括黏附损伤、膜融合损伤、入侵伤害、毒性损伤、免疫损伤和炎症损伤，对实验室常用的分子诊断技术进行论述，涵盖核酸恒温扩增技术与变温扩增技术，恒温扩增技术例如环介导等温扩增技术（LAMP）、链置换反应、多重依赖核酸序列的扩增技术（NASBA）、重组酶扩增技术（RAA），变温扩增技术例如传统聚合酶链式反应（PCR）、广泛PCR、巢式PCR、实时PCR、多重PCR，并在检测技术基础上综合论述了肺炎支原体检测的生物传感平台，囊括侧流层析传感器、电化学传感器、荧光传感器等。旨在总结目前针对肺炎支原体检测技术的优缺点，以期为肺炎支原体的早期诊断提供一定的方法参考，展望未来肺炎支原体免提取、一体化、快速检测，伴随检测成本的降低，使得患者可以实现居家自检，避免抗感染药物的滥用，迎接临床诊疗进入精准时代。

关键词: 肺炎支原体, 核酸分子诊断, 恒温扩增, 变温扩增, 生物传感器

Abstract:

Mycoplasma pneumoniae is the smallest known cell organism, and is one of major pathogens causing community-acquired pneumonia. The symptoms of infection can differ widely in the early stages and can affect different organs in a body. Early diagnosis is challenging because early clinical symptoms and radiological examinations lack specificity and are prone to misdiagnosis and missed diagnosis, which can threaten physical health. At present, both domestic and international M. pneumoniae detection mainly relies on laboratory diagnostic methods. This article begins by examining the intricate pathogenesis of M. pneumoniae, which includes adhesion damage, membrane fusion damage, invasion damage, toxicity damage, immune damage, and inflammatory damage. It also briefly explores commonly used molecular diagnostic techniques in the laboratory, such as nucleic acid isothermal amplification technology and variable temperature amplification technology. The isothermal amplification techniques include loop-mediated isothermal amplification（LAMP）, chain displacement reaction, sequence-dependent nucleic acid amplification（NASBA）, and recombinase-aided amplification（RAA）. Additionally, variable temperature amplification techniques refer to traditional polymerase chain reaction（PCR）, broad-range PCR, nested PCR, real-time PCR, and multiplex PCR. The text also includes a comprehensive discussion on biosensing platforms for M. pneumoniae detection, which encompasses lateral flow assay, electrochemical biosensors, fluorescence biosensors, and more. This manuscript aims to summarize the advantages and disadvantages of current M. pneumoniae detection techniques, providing a reference for early diagnosis. It also anticipates future non-extraction, integration, and rapid detection methods. Along with reduced costs, these advancements could enable patients to perform self-examinations at home, avoid unnecessary use of anti-infective drugs, and usher in a precision era for clinical diagnosis and treatment.

Key words: Mycoplasma pneumoniae, nucleic acid molecular diagnosis, isothermal amplification, variable temperature amplification, biosensor

余永霞, 祝宁, 刘光敏, 朱龙佼, 许文涛. 肺炎支原体核酸分子诊断技术研究进展[J]. 生物技术通报, 2024, 40(12): 72-83.

YU Yong-xia, ZHU Ning, LIU Guang-min, ZHU Long-jiao, XU Wen-tao. Research Progress in Nucleic Acid Molecular Diagnostic Technology for Mycoplasma pneumoniae[J]. Biotechnology Bulletin, 2024, 40(12): 72-83.

图/表 4

参考文献 74

[1]	Waites KB, Talkington DF. Mycoplasma pneumoniaeand its role as a human pathogen[J]. Clin Microbiol Rev, 2004, 17(4): 697-728. pmid: 15489344
[2]	Chanock RM, Hayflick L, Barile MF. Growth on artificial medium of an agent associated with atypical pneumonia and its identification as a PPLO[J]. Proc Natl Acad Sci U S A, 1962, 48(1): 41-49.
[3]	Kutty PK, Jain S, Taylor TH, et al. Mycoplasma pneumoniae among children hospitalized with community-acquired pneumonia[J]. Clin Infect Dis, 2019, 68(1): 5-12. doi: 10.1093/cid/ciy419 pmid: 29788037
[4]	Smith CB, Chanock RM, Friedewald WT, et al. Mycoplasma pneumoniae infections in volunteers[J]. Ann N Y Acad Sci, 1967, 143(1): 471-483.
[5]	孙天文, 刘思辰, 杨柯. 实时荧光核酸恒温扩增试验对肺炎支原体感染的诊断价值[J]. 河南医学高等专科学校学报, 2023, 35(5): 543-547.
	Sun TW, Liu SC, Yang K. Diagnostic value of real-time fluorescent nucleic acid thermostatic amplification test for Mycoplasma pneumoniae infection[J]. J Henan Med Coll, 2023, 35(5): 543-547.
[6]	Shimizu T, Kimura Y, Kida Y, et al. Cytadherence of Mycoplasma pneumoniae induces inflammatory responses through autophagy and toll-like receptor 4[J]. Infect Immun, 2014, 82(7): 3076-3086. doi: 10.1128/IAI.01961-14 pmid: 24799628
[7]	Chourasia BK, Chaudhry R, Malhotra P. Delineation of immunodominant and cytadherence segment(s)of Mycoplasma pneumoniae P1 gene[J]. BMC Microbiol, 2014, 14: 108.
[8]	Seto S, Kenri T, Tomiyama T, et al. Involvement of P1 adhesin in gliding motility of Mycoplasma pneumoniae as revealed by the inhibitory effects of antibody under optimized gliding conditions[J]. J Bacteriol, 2005, 187(5): 1875-1877. pmid: 15716461
[9]	Balish MF, Santurri RT, Ricci AM, et al. Localization of Mycoplasma pneumoniae cytadherence-associated protein HMW2 by fusion with green fluorescent protein: implications for attachment organelle structure[J]. Mol Microbiol, 2003, 47(1): 49-60. pmid: 12492853
[10]	McDermott AJ, Taylor BM, Bernstein KM. Toxic epidermal necrolysis from suspected Mycoplasma pneumoniae infection[J]. Mil Med, 2013, 178(9): e1048-e1050.
[11]	Ledford JG, Mukherjee S, Kislan MM, et al. Surfactant protein-a suppresses eosinophil-mediated killing of Mycoplasma pneumoniae in allergic lungs[J]. PLoS One, 2012, 7(2): e32436.
[12]	Lai JF, Zindl CL, Duffy LB, et al. Critical role of macrophages and their activation via MyD88-NFκB signaling in lung innate immunity to Mycoplasma pneumoniae[J]. PLoS One, 2010, 5(12): e14417.
[13]	庞焕香, 乔红梅, 成焕吉, 等. 支原体肺炎患儿肺泡灌洗液中TNF-α、IL-6、IL-10水平检测及意义[J]. 中国当代儿科杂志, 2011, 13(10): 808-810.
	Pang HX, Qiao HM, Cheng HJ, et al. Levels of TNF-α, IL-6 and IL-10 in bronchoalveolar lavage fluid in children with Mycoplasma pneumoniae pneumonia[J]. Chin J Contemp Pediatr, 2011, 13(10): 808-810.
[14]	中华医学会儿科学分会临床检验学组. 儿童肺炎支原体呼吸道感染实验室诊断中国专家共识[J]. 中华检验医学杂志, 2019, 42(7): 507-513.
	Clinical Laboratory Group, Pediatrics Society of Chinese Medical Association. Chinese expert consensus on laboratory diagnosis of mycoplasma pneumoniae respiratory tract infection in children[J]. Chin J Lab Med, 2019, 42(7): 507-513.
[15]	Ieven M, Ursi D, Van Bever H, et al. Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of M. pneumoniae in acute respiratory tract infections in pediatric patients[J]. J Infect Dis, 1996, 173(6): 1445-1452. pmid: 8648218
[16]	王居鹏, 朱黎娜, 马明坤, 等. 被动凝集法、间接免疫荧光法和胶体金法联合检测肺炎支原体抗体对儿童肺炎支原体感染的诊断价值[J]. 天津医药, 2022, 50(4): 418-423.
	Wang JP, Zhu LN, Ma MK, et al. Diagnostic value of particle agglutination, indirect immunofluorescence assay and immune colloidal gold technique combined detection for Mycoplasma pneumoniae antibody in children with Mycoplasma pneumoniae infection[J]. Tianjin Med J, 2022, 50(4): 418-423.
[17]	Waites KB, Xiao L, Paralanov V, et al. Molecular methods for the detection of Mycoplasma and ureaplasma infections in humans: a paper from the 2011 William Beaumont Hospital Symposium on molecular pathology[J]. J Mol Diagn, 2012, 14(5): 437-450. doi: 10.1016/j.jmoldx.2012.06.001 pmid: 22819362
[18]	Bernet C, Garret M, de Barbeyrac B, et al. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction[J]. J Clin Microbiol, 1989, 27(11): 2492-2496. doi: 10.1128/jcm.27.11.2492-2496.1989 pmid: 2509513
[19]	Dorigo-Zetsma JW, Verkooyen RP, van Helden HP, et al. Molecular detection of Mycoplasma pneumoniae in adults with community-acquired pneumonia requiring hospitalization[J]. J Clin Microbiol, 2001, 39(3): 1184-1186. pmid: 11230455
[20]	Templeton KE, Scheltinga SA, Graffelman AW, et al. Comparison and evaluation of real-time PCR, real-time nucleic acid sequence-based amplification, conventional PCR, and serology for diagnosis of Mycoplasma pneumoniae[J]. J Clin Microbiol, 2003, 41(9): 4366-4371. doi: 10.1128/JCM.41.9.4366-4371.2003 pmid: 12958270
[21]	Medjo B, Atanaskovic-Markovic M, Radic S, et al. Mycoplasma pneumoniae as a causative agent of community-acquired pneumonia in children: clinical features and laboratory diagnosis[J]. Ital J Pediatr, 2014, 40: 104. doi: 10.1186/s13052-014-0104-4 pmid: 25518734
[22]	Morozumi M, Hasegawa K, Chiba N, et al. Application of PCR for Mycoplasma pneumoniae detection in children with community-acquired pneumonia[J]. J Infect Chemother, 2004, 10(5): 274-279. pmid: 16163461
[23]	Roth SB, Jalava J, Ruuskanen O, et al. Use of an oligonucleotide array for laboratory diagnosis of bacteria responsible for acute upper respiratory infections[J]. J Clin Microbiol, 2004, 42(9): 4268-4274. pmid: 15365022
[24]	Störmer M, Vollmer T, Henrich B, et al. Broad-range real-time PCR assay for the rapid identification of cell-line contaminants and clinically important mollicute species[J]. Int J Med Microbiol, 2009, 299(4): 291-300. doi: 10.1016/j.ijmm.2008.08.002 pmid: 18926769
[25]	Wang H, Kong FR, Jelfs P, et al. Simultaneous detection and identification of common cell culture contaminant and pathogenic mollicutes strains by reverse line blot hybridization[J]. Appl Environ Microbiol, 2004, 70(3): 1483-1486.
[26]	Lin BC, Blaney KM, Malanoski AP, et al. Using a resequencing microarray as a multiple respiratory pathogen detection assay[J]. J Clin Microbiol, 2007, 45(2): 443-452. pmid: 17135438
[27]	Kumar S, Bharti PK, Baveja CP, et al. Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of Mycoplasma pneumoniae in pediatric community-acquired lower respiratory tract infections[J]. Indian J Med Microbiol, 2022, 40(2): 250-253.
[28]	Guo DX, Hu WJ, Wei R, et al. Epidemiology and mechanism of drug resistance of Mycoplasma pneumoniae in Beijing, China: a multicenter study[J]. Bosn J Basic Med Sci, 2019, 19(3): 288-296.
[29]	Raggam RB, Leitner E, Berg J, et al. Single-Run, parallel detection of DNA from three pneumonia-producing bacteria by real-time polymerase chain reaction[J]. J Mol Diagn, 2005, 7(1): 133-138. pmid: 15681485
[30]	Li LF, Ma JY, Guo PB, et al. Molecular beacon based real-time PCR p1 gene genotyping, macrolide resistance mutation detection and clinical characteristics analysis of Mycoplasma pneumoniae infections in children[J]. BMC Infect Dis, 2022, 22(1): 724.
[31]	Pitcher D, Chalker VJ, Sheppard C, et al. Real-time detection of Mycoplasma pneumoniae in respiratory samples with an internal processing control[J]. J Med Microbiol, 2006, 55(Pt 2): 149-155. doi: 10.1099/jmm.0.46281-0 pmid: 16434706
[32]	Dumke R, Schurwanz N, Lenz M, et al. Sensitive detection of Mycoplasma pneumoniae in human respiratory tract samples by optimized real-time PCR approach[J]. J Clin Microbiol, 2007, 45(8): 2726-2730. pmid: 17537933
[33]	Winchell JM, Thurman KA, Mitchell SL, et al. Evaluation of three real-time PCR assays for detection of Mycoplasma pneumoniae in an outbreak investigation[J]. J Clin Microbiol, 2008, 46(9): 3116-3118. doi: 10.1128/JCM.00440-08 pmid: 18614663
[34]	Liu Y, Ye XY, Zhang H, et al. Rapid detection of Mycoplasma pneumoniae and its macrolide-resistance mutation by Cycleave PCR[J]. Diagn Microbiol Infect Dis, 2014, 78(4): 333-337.
[35]	Tang MY, Wang D, Tong X, et al. Comparison of different detection methods for Mycoplasma pneumoniae infection in children with community-acquired pneumonia[J]. BMC Pediatr, 2021, 21(1): 90.
[36]	Wang JC, Guo CY, Yang LX, et al. Peripheral blood microR-146a and microR-29c expression in children with Mycoplasma pneumoniae pneumonia and its clinical value[J]. Ital J Pediatr, 2023, 49(1): 119.
[37]	Ji M, Lee NS, Oh JM, et al. Single-nucleotide polymorphism PCR for the detection of Mycoplasma pneumoniae and determination of macrolide resistance in respiratory samples[J]. J Microbiol Methods, 2014, 102: 32-36. doi: 10.1016/j.mimet.2014.04.009 pmid: 24780151
[38]	Zacharioudakis IM, Zervou FN, Dubrovskaya Y, et al. Evaluation of a multiplex PCR panel for the microbiological diagnosis of pneumonia in hospitalized patients: experience from an academic medical center[J]. Int J Infect Dis, 2021, 104: 354-360. doi: 10.1016/j.ijid.2021.01.004 pmid: 33434669
[39]	Miyashita N, Saito A, Kohno S, et al. Multiplex PCR for the simultaneous detection of Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila in community-acquired pneumonia[J]. Respir Med, 2004, 98(6): 542-550.
[40]	Geertsen R, Kaeppeli F, Sterk-Kuzmanovic N, et al. A multiplex PCR assay for the detection of respiratory bacteriae in nasopharyngeal smears from children with acute respiratory disease[J]. Scand J Infect Dis, 2007, 39(9): 769-774. pmid: 17701714
[41]	Kumar S, Wang LH, Fan J, et al. Detection of 11 common viral and bacterial pathogens causing community-acquired pneumonia or sepsis in asymptomatic patients by using a multiplex reverse transcription-PCR assay with manual(enzyme hybridization)or automated(electronic microarray)detection[J]. J Clin Microbiol, 2008, 46(9): 3063-3072.
[42]	Lodes MJ, Suciu D, Wilmoth JL, et al. Identification of upper respiratory tract pathogens using electrochemical detection on an oligonucleotide microarray[J]. PLoS One, 2007, 2(9): e924.
[43]	Benson R, Tondella ML, Bhatnagar J, et al. Development and evaluation of a novel multiplex PCR technology for molecular differential detection of bacterial respiratory disease pathogens[J]. J Clin Microbiol, 2008, 46(6): 2074-2077. doi: 10.1128/JCM.01858-07 pmid: 18400916
[44]	Wang YJ, Kong FR, Yang YH, et al. A multiplex PCR-based reverse line blot hybridization(mPCR/RLB)assay for detection of bacterial respiratory pathogens in children with pneumonia[J]. Pediatr Pulmonol, 2008, 43(2): 150-159.
[45]	Shen HW, Zhu BQ, Wang SL, et al. Association of targeted multiplex PCR with resequencing microarray for the detection of multiple respiratory pathogens[J]. Front Microbiol, 2015, 6: 532. doi: 10.3389/fmicb.2015.00532 pmid: 26074910
[46]	Wang L, Feng ZS, Zhao MC, et al. A comparison study between GeXP-based multiplex-PCR and serology assay for Mycoplasma pneumoniae detection in children with community acquired pneumonia[J]. BMC Infect Dis, 2017, 17(1): 518.
[47]	Loens K, Van Heirstraeten L, Malhotra-Kumar S, et al. Optimal sampling sites and methods for detection of pathogens possibly causing community-acquired lower respiratory tract infections[J]. J Clin Microbiol, 2009, 47(1): 21-31. doi: 10.1128/JCM.02037-08 pmid: 19020070
[48]	Diaz MH, Winchell JM. Detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae directly from respiratory clinical specimens using a rapid real-time polymerase chain reaction assay[J]. Diagn Microbiol Infect Dis, 2012, 73(3): 278-280.
[49]	Pillet S, Lardeux M, Dina JL, et al. Comparative evaluation of six commercialized multiplex PCR kits for the diagnosis of respiratory infections[J]. PLoS One, 2013, 8(8): e72174.
[50]	Notomi T, Okayama H, Masubuchi H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res, 2000, 28(12): E63.
[51]	Petrone BL, Wolff BJ, DeLaney AA, et al. Isothermal detection of Mycoplasma pneumoniae directly from respiratory clinical specimens[J]. J Clin Microbiol, 2015, 53(9): 2970-2976. doi: 10.1128/JCM.01431-15 pmid: 26179304
[52]	Ratliff AE, Duffy LB, Waites KB. Comparison of the illumigene Mycoplasma DNA amplification assay and culture for detection of Mycoplasma pneumoniae[J]. J Clin Microbiol, 2014, 52(4): 1060-1063. doi: 10.1128/JCM.02913-13 pmid: 24430454
[53]	Shi C, Shang FJ, Zhou ML, et al. Triggered isothermal PCR by denaturation bubble-mediated strand exchange amplification[J]. Chem Commun, 2016, 52(77): 11551-11554.
[54]	Yang C, Li Y, Deng J, et al. Accurate, rapid and low-cost diagnosis of Mycoplasma pneumoniae via fast narrow-thermal-cycling denaturation bubble-mediated strand exchange amplification[J]. Anal Bioanal Chem, 2020, 412(30): 8391-8399. doi: 10.1007/s00216-020-02977-y pmid: 33040157
[55]	Diaz MH, Winchell JM. The evolution of advanced molecular diagnostics for the detection and characterization of Mycoplasma pneumoniae[J]. Front Microbiol, 2016, 7: 232. doi: 10.3389/fmicb.2016.00232 pmid: 27014191
[56]	Loens K, Ieven M, Ursi D, et al. Application of NucliSens Basic Kit for the detection of Mycoplasma pneumoniae in respiratory specimens[J]. J Microbiol Methods, 2003, 54(1): 127-130. pmid: 12732431
[57]	Loens K, Beck T, Ursi D, et al. Development of real-time multiplex nucleic acid sequence-based amplification for detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella spp. in respiratory specimens[J]. J Clin Microbiol, 2008, 46(1): 185-191. doi: 10.1128/JCM.00447-07 pmid: 18032625
[58]	Jiang TT, Wang YC, Jiao WW, et al. Recombinase polymerase amplification combined with real-time fluorescent probe for Mycoplasma pneumoniae detection[J]. J Clin Med, 2022, 11(7): 1780.
[59]	Zhou J, Xiao F, Fu J, et al. Rapid, ultrasensitive and highly specific diagnosis of Mycoplasma pneumoniae by a CRISPR-based detection platform[J]. Front Cell Infect Microbiol, 2023, 13: 1147142.
[60]	Xue GH, Li SL, Zhao HQ, et al. Use of a rapid recombinase-aided amplification assay for Mycoplasma pneumoniae detection[J]. BMC Infect Dis, 2020, 20(1): 79.
[61]	Xia SM, Chen X. Single-copy sensitive, field-deployable, and simultaneous dual-gene detection of SARS-CoV-2 RNA via modified RT-RPA[J]. Cell Discov, 2020, 6(1): 37.
[62]	Deng ZL, Hu HY, Tang D, et al. Ultrasensitive, specific, and rapid detection of Mycoplasma pneumoniae using the ERA/CRISPR-Cas12a dual system[J]. Front Microbiol, 2022, 13: 811768.
[63]	Wang YC, Wang Y, Quan ST, et al. Establishment and application of a multiple cross displacement amplification coupled with nanoparticle-based lateral flow biosensor assay for detection of Mycoplasma pneumoniae[J]. Front Cell Infect Microbiol, 2019, 9: 325.
[64]	Zhu MJ, Ma L, Meng QF, et al. Visual detection of Mycoplasma pneumoniae by the recombinase polymerase amplification assay coupled with lateral flow dipstick[J]. J Microbiol Methods, 2022, 202: 106591.
[65]	Hu LY, Wang XR, Cao DL, et al. Establishment and performance evaluation of multiplex PCR-dipstick DNA chromatography for Mycoplasma pneumoniae and Chlamydia pneumoniae rapid detection[J]. Can J Infect Dis Med Microbiol, 2023, 2023: 6654504.
[66]	Yang W, Restrepo-Pérez L, Bengtson M, et al. Detection of CRISPR-dCas9 on DNA with solid-state nanopores[J]. Nano Lett, 2018, 18(10): 6469-6474. doi: 10.1021/acs.nanolett.8b02968 pmid: 30187755
[67]	Zhu R, Jiang H, Li CY, et al. CRISPR/Cas9-based point-of-care lateral flow biosensor with improved performance for rapid and robust detection of Mycoplasma pneumonia[J]. Anal Chim Acta, 2023, 1257: 341175.
[68]	Liu LL, Xiang GM, Jiang DN, et al. Electrochemical gene sensor for Mycoplasma pneumoniae DNA using dual signal amplification via a Pt@Pd nanowire and horse radish peroxidase[J]. Microchim Acta, 2016, 183(1): 379-387.
[69]	Sen D, Gilbert W. A sodium-potassium switch in the formation of four-stranded G4-DNA[J]. Nature, 1990, 344(6265): 410-414.
[70]	Li JJ, Wu J, He ZQ, et al. Fast detection of Mycoplasma pneumoniae by interaction of tetramolecular G-quadruplex with graphene oxide[J]. Sens Actuat B Chem, 2019, 290: 41-46.
[71]	Wang H, Ma Z, Qin JX, et al. A versatile loop-mediated isothermal amplification microchip platform for Streptococcus pneumoniae and Mycoplasma pneumoniae testing at the point of care[J]. Biosens Bioelectron, 2019, 126: 373-380. doi: S0956-5663(18)30905-9 pmid: 30469075
[72]	Mongan AE, Tuda JSB, Runtuwene LR. Portable sequencer in the fight against infectious disease[J]. J Hum Genet, 2020, 65(1): 35-40. doi: 10.1038/s10038-019-0675-4 pmid: 31582773
[73]	Ishiguro N, Sato R, Mori T, et al. Point-of-care molecular diagnosis of Mycoplasma pneumoniae including macrolide sensitivity using quenching probe polymerase chain reaction[J]. PLoS One, 2021, 16(10): e0258694.
[74]	Duan YK, Zhang X, Li Y, et al. Amino-modified silica membrane capable of DNA extraction and enrichment for facilitated isothermal amplification detection of Mycoplasma pneumoniae[J]. J Pharm Biomed Anal, 2023, 224: 115190.

编辑推荐 0

Metrics

阅读次数

全文

HTML			PDF

最新录用	在线预览	正式出版	最新录用	在线预览	正式出版
0	0	6	0	0	48

来源	本网站	其他网站

次数	47	7
比例	87%	13%

摘要

最新录用	在线预览	正式出版

0	0	88

	来源	本网站

	次数	88
	比例	100%

方法Method	靶标位点Target site	使用样本Sample type	参考文献Reference
传统PCR Traditional PCR	P1黏附素基因	血液/咽拭子	[20-21]
传统PCR Traditional PCR	16S rRNA	鼻咽分泌物/咽拭子/痰液	[22]
广泛 PCR Broad-range PCR	part E	咽拭子	[23]
广泛 PCR Broad-range PCR	tuf	咽拭子/痰液	[24]
巢式PCR Nested PCR	16S-23S rRNA 间隔区	鼻咽分泌物	[25]
	P1黏附素基因	咽拭子/鼻咽分泌物	[26-27]
	23S rRNA	气管拭子	[28]
实时PCR Real-time PCR	16S rRNA	痰液/咽拭子/肺泡灌洗液	[29-30]
	P1黏附素基因	痰液	[31]
	repMp1	支气管肺泡灌洗液/鼻咽拭子	[32]
	CARDS/ATPase	鼻咽拭子	[33]
	23S rRNA	鼻咽拭子/痰液	[34-35]
	microR-146/cmicroR-17a	外周血/痰液	[36]
多重PCR Multiplex PCR	23S rRNA	痰液/肺泡灌洗液	[37-38]
	16S rRNA	鼻咽拭子	[39]
	P1黏附素基因	鼻咽拭子/血液	[40-41]
	dnak/pdhA/tuf	ATCC标准品	[42]
	ATPase	鼻咽拭子	[43]
	16S-23S rRNA 间隔区	鼻咽拭子	[44]
	CARDS	鼻咽分泌物	[45]
	16S rRNA	痰液	[46]

方法Method	靶标位点Target site	使用样本Sample type	参考文献Reference
传统PCR Traditional PCR	P1黏附素基因	血液/咽拭子	[20-21]
传统PCR Traditional PCR	16S rRNA	鼻咽分泌物/咽拭子/痰液	[22]
广泛 PCR Broad-range PCR	part E	咽拭子	[23]
广泛 PCR Broad-range PCR	tuf	咽拭子/痰液	[24]
巢式PCR Nested PCR	16S-23S rRNA 间隔区	鼻咽分泌物	[25]
	P1黏附素基因	咽拭子/鼻咽分泌物	[26-27]
	23S rRNA	气管拭子	[28]
实时PCR Real-time PCR	16S rRNA	痰液/咽拭子/肺泡灌洗液	[29-30]
	P1黏附素基因	痰液	[31]
	repMp1	支气管肺泡灌洗液/鼻咽拭子	[32]
	CARDS/ATPase	鼻咽拭子	[33]
	23S rRNA	鼻咽拭子/痰液	[34-35]
	microR-146/cmicroR-17a	外周血/痰液	[36]
多重PCR Multiplex PCR	23S rRNA	痰液/肺泡灌洗液	[37-38]
	16S rRNA	鼻咽拭子	[39]
	P1黏附素基因	鼻咽拭子/血液	[40-41]
	dnak/pdhA/tuf	ATCC标准品	[42]
	ATPase	鼻咽拭子	[43]
	16S-23S rRNA 间隔区	鼻咽拭子	[44]
	CARDS	鼻咽分泌物	[45]
	16S rRNA	痰液	[46]