血流感染分子诊断的研究进展

doi:10.3969/j.issn.1673-8640.2014.06.002

摘要/Abstract

摘要：

血流感染是一种严重的全身感染性疾病,血培养仍是目前诊断细菌性血流感染的金标准,但仅有30％～40%的血流感染可通过培养发现致病菌。分子生物学方法可通过分析患者血液标本中病原微生物的核酸成分,快速提供准确的细菌、真菌或病毒感染信息,甚至提供常见病原菌的耐药基因检测结果。目前,应用于临床实验室血流感染检测的分子生物学技术主要包括核酸杂交技术、核酸扩增及DNA序列分析、基因芯片和质谱检测技术等。对常见致病菌、分枝杆菌、苛养菌、少见病原菌等使用多种检测技术联合应用进行快速鉴定及耐药分析,可以在较短时间内为临床提供可靠的诊断结果,提示临床合理用药,提高血流感染患者的存活率。

关键词: 血流感染, 分子诊断, 研究进展

Abstract:

The blood stream infection is a syndrome of serious systemic infection, and blood culture remains being the gold standard for the diagnosis of bacterial blood stream infection, but only 30%-40% blood stream infection pathogens could be found by blood culture. Molecular biology method through analyzing the nucleic acid of microorganisms in blood samples will quickly provide the results of bacterial, fungal or viral infections accurately, and provide a common resistance gene detection results of pathogens. At present, the molecular diagnosis techniques for blood stream infection in clinical laboratoriesmainly include nucleic acid hybridization, nucleic acid amplification and DNA sequence analysis, gene chip, mass spectrometry and so on. The common pathogenic bacteria, mycobacteria, fastidious bacteria, rare pathogenic bacteria use a variety of detection techniques combined with the application of rapid identification and drug resistance analysis, which can provide reliable diagnosis results for clinical in a relatively short period of time, indicate the use of clinical rational drug and improve survival in patients with blood stream infection.

Key words: Blood stream infection, Molecular diagnosis, Research progress

中图分类号:

R446.5

郭建综述,吴文娟审校. 血流感染分子诊断的研究进展[J]. 检验医学, 2014, 29(6): 584-589.

GUO Jian,WU Wenjuan. The research progress in the molecular diagnosis of blood stream infection[J]. , 2014, 29(6): 584-589.

参考文献

[1] Skvarc M, Stubljar D, Rogina P, et al. Non-culture-based methods to diagnose bloodstream infection: does it work[J] . Eur J Microbiol Immunol (Bp),2013, 3(2):97-104.
[2] Afshari A, Schrenzel J, Ieven M, et al. Bench-to-bedside review: rapid molecular diagnostics for bloodstream infection-a new frontierr[J] . Crit Care, 2012,16(3):222.
[3] Loonen AJ, de Jager CP, Tosserams J, et al. Biomarkers and molecular analysis to improve bloodstream infection diagnostics in an emergency care unit[J] . PLoS One, 2014,9(1):e87315.
[4] Deck MK, Anderson ES, Buckner RJ, et al. Multicenter evaluation of the Staphylococcus QuickFISH method for simultaneous identification of Staphylococcus aureus and coagulase-negative staphylococci directly from blood culture bottles in less than 30 minutes[J] . J Clin Microbiol, 2012,50(6):1994-1998.
[5] Forrest GN, Roghmann MC, Toombs LS, et al. Peptide nucleic acid fluorescent in situ hybridization for hospital-acquired enterococcal bacteremia: delivering earlier effective antimicrobial therapy[J] . Antimicrob Agents Chemother, 2008,52(10):3558-3563.
[6] Forrest GN, Mehta S, Weekes E, et al. Impact of rapid in situ hybridization testing on coagulase-negative staphylococci positive blood cultures[J] . J Antimicrob Chemother,2006,58(1):154-158.
[7] Pasko C, Hicke B, Dunn J, et al. Staph ID/R: a rapid method for determining staphylococcus species identity and detecting the mecA gene directly from positive blood culture[J] . J Clin Microbiol, 2012,50(3):810-817.
[8] Sidor IF, Dunn JL, Tsongalis GJ, et al. A multiplex real-time polymerase chain reaction assay with two internal controls for the detection of Brucella species in tissues, blood, and feces from marine mammals[J] . J Vet Diagn Invest, 2013,25(1):72-81.
[9] Springer J, Morton CO, Perry M, et al. Multicenter comparison of serum and whole-blood specimens for detection of Aspergillus DNA in high-risk hematological patients[J] . J Clin Microbiol,2013,51(5):1445-1450.
[10] Karah N, Haldorsen B, Hegstad K, et al. Species identification and molecular characterization of Acinetobacter spp. blood culture isolates from Norway[J] .J Antimicrob Chemother,2011, 66(4): 738-744.
[11] Benítez-Páez A, lvarez M, Belda-Ferre P, et al. Detection of transient bacteraemia following dental extractions by 16S rDNA pyrosequencing: a pilot study[J] . PLoS One,2013,8(3):e57782.
[12] Jeng K, Yang S, Won H, et al. Application of a 16S rRNA PCR-high-resolution melt analysis assay for rapid detection of Salmonella Bacteremia[J] . J Clin Microbiol, 2012,50(3):1122-1124.
[13] Steindor M, Weizenegger M, Harrison N, et al. Use of a commercial PCR-based line blot method for identification of bacterial pathogens and the mecA and van genes from BacTAlert blood culture bottles[J] . J Clin Microbiol, 2012,50(1):157-159.
[14] Hansen WL, Beuving J, Verbon A, et al. One-day workflow scheme for bacterial pathogen detection and antimicrobial resistance testing from blood cultures[J] . J Vis Exp,2012,9(65): 3254.
[15] Dark P, Wilson C, Blackwood B. Accuracy of LightCycler(R) SeptiFast for the detection and identification of pathogens in the blood of patients with suspected sepsis: a systematic review protocol[J] . BMJ Open, 2012, 2(1):e000392.
[16] Laakso S, Mki M. Assessment of a semi-automated protocol for multiplex analysis of sepsis-causing bacteria with spiked whole blood samples[J] . Microbiologyopen,2013, 2(2):284-292.
[17] Lucignano B, Ranno S, Liesenfeld O, et al. Multiplex PCR allows rapid and accurate diagnosis of bloodstream infections in newborns and children with suspected sepsis[J] . J Clin Microbiol,2011,49(6):2252-2258.
[18] Wojewoda CM, Sercia L, Navas M, et al. Evaluation of the Verigene Gram-positive blood culture nucleic acid test for rapid detection of bacteria and resistance determinants[J] . J Clin Microbiol, 2013,51(7):2072-2076.
[19] Pichon B, Hill R, Laurent F, et al. Development of a real-time quadruplex PCR assay for simultaneous detection of nuc, Panton-Valentine leucocidin (PVL), mecA and homologue mecALGA251[J] . J Antimicrob Chemother,2012, 67(10):2338-2341.
[20] Wang HY, Kim S, Kim H, et al. Real-time PCR TaqMan assay for rapid screening of bloodstream infection[J] . Ann Clin Microbiol Antimicrob,2014,13(1):3.
[21] Samuel LP, Tibbetts RJ, Agotesku A, et al. Evaluation of a microarray-based assay for rapid identification of Gram-positive organisms and resistance markers in positive blood cultures[J] . J Clin Microbiol,2013,51(4):1188-1192.
[22] Schubert S, Weinert K, Wagner C, et al. Novel, improved sample preparation for rapid, direct identification from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry[J] . J Mol Diagn,2011,13(6):701-706.
[23] Jeng K, Gaydos CA, Blyn LB, et al. Comparative analysis of two broad-range PCR assays for pathogen detection in positive-blood-culture bottles: PCR-high-resolution melting analysis versus PCR-mass spectrometry[J] . J Clin Microbiol,2012,50(10):3287-3292.
[24] Jordana-Lluch E, Carolan HE, Giménez M, et al. Rapid diagnosis of bloodstream infections with PCR followed by mass spectrometry[J] . PLoS One,2013,8(4):e62108.
[25] Kok J, Thomas LC, Olma T, et al. Identification of bacteria in blood culture broths using matrix-assisted laser desorption-ionization Sepsityper^TM and time of flight mass spectrometry[J] . PLoS One, 2011,6(8):e23285.
[26] Feasey NA, Banada PP, Howson W, et al. Evaluation of Xpert MTB/RIF for detection of tuberculosis from blood samples of HIV-infected adults confirms Mycobacterium tuberculosis bacteremia as an indicator of poor prognosis[J] . J Clin Microbiol,2013,51(7):2311-2316.
[27] Banada PP, Koshy R, Alland D. Detection of Mycobacterium tuberculosis in blood by use of the Xpert MTB/RIF assay[J] . J Clin Microbiol,2013,51(7):2317-2322.
[28] Fu Y, Yi Z, Wu X, et al. Circulating microRNAs in patients with active pulmonary tuberculosis[J] . J Clin Microbiol, 2011,49(12):4246-4251.