双歧杆菌制剂辅助治疗儿童抗生素相关性腹泻的系统评价与Meta分析

目的评价双歧杆菌制剂辅助治疗儿童抗生素相关性腹泻的临床疗效。方法检索CNKI、维普、万方、PubMed、Embase等数据库,时间为建库至2019年8月,筛选随机对照试验(randomized controlled tests,RCTs)。采用RevMan 5.3软件对纳入的研究进行Meta分析,采用Cochrane系统评价工具评估文献发表偏倚。结果共纳入7篇较高质量的RCTs,Meta分析结果示:RR=1.21,95%CI=(1.14,1.27),Z=6.80,P0.01。结论双歧杆菌制剂可有效提高患者的总有效率,改善患者症状。但纳入文献存在一定的发表偏倚,仍需要更多的研究数据加以验证。     

双歧杆菌制剂对肿瘤化疗患者肠道菌群影响的Meta分析

目的 探讨双歧杆菌制剂对肿瘤化疗患者肠道菌群的影响。方法 通过检索PubMed、Embase、The Cochrane Library、Web of Science以及CBM、CNKI、WanFang Data和VIP数据库,收集各数据库从建库至2022年7月发表的所有关于肿瘤化疗患者应用双歧杆菌制剂的随机对照试验,由2名研究者独立筛选文献,按照系统评价的要求对文献质量进行评估,使用Revman 5.4和Stata 17.0统计学软件进行Meta分析。结果 共纳入25篇文献（2 152例患者）,Meta分析结果显示,双歧杆菌制剂组患者肠道中双歧杆菌和乳杆菌的数量高于对照组[SMD=2.89, 95%CI(2.28, 3.49), P<0.001; SMD=2.20, 95%CI(1.57, 2.84),P<0.001],双歧杆菌制剂组患者肠道中大肠埃希菌、肠杆菌和肠球菌的数量低于对照组[SMD=-1.08,95%CI（-1.64,-0.52）, P<0.001; SMD=-0.88, 95%CI（-1.58,-0.17）, P=0.010; SMD=-0.98, 95...     

荧光定量PCR检测结核分枝杆菌Meta分析

目的系统评价荧光定量PCR(FQ-PCR)方法检测结核分枝杆菌的效果。方法按照系统评价的要求检索CBM、VIP、CNKI以及万方数据库等,获得20篇符合纳入标准的文献,对其进行Meta分析,并评价Meta分析结果的稳定性和发表偏倚。结果 FQ-PCR对照涂片染色、培养鉴定以及总数据的异质性检验P0.00001,采用随机效应模型进行Meta分析,其余的采用固定效应模型分析。FQ-PCR与涂片染色、培养鉴定、抗体检测等的总体效应Z值分别为7.76、5.00和7.34,P值均小于0.00001,差异具有统计学意义。总数据分析结果的合并OR=2.78,95%CI为1.93-4.01,总体效应检验,Z=5.49,P0.00001,差异具有统计学意义,固定效应模型OR值和95%CI(2.52[2.35-2.70])与随机效应模型比较接近,剔除小样本报道后的合并OR=2.93,95%CI为1.98-4.31,与剔除前的结果也比较接近。结论从现有的临床证据来看,FQ-PCR是检测结核分枝杆菌的有效方法,可推广应用与临床结核病辅助检测。     

阴道用乳酸菌制剂辅助治疗RVVC的Meta分析

目的系统评价阴道用乳酸菌制剂辅助治疗复发性外阴阴道假丝酵母菌病(RVVC)的疗效及安全性。方法计算机检索CNKI、万方、维普、PubMed及Cochrane等数据库(检索时间为各数据库创建时间至2018年1月),纳入关于阴道用乳酸菌制剂辅助治疗RVVC的临床随机对照试验(RCT),采用RevMan 5.3软件对相关研究进行Meta分析。结果共纳入32篇RCT,包括3 635例患者,结果显示:阴道用乳酸菌制剂辅助治疗RVVC的痊愈率优于单纯抗菌药物[P0.001,OR=2.15,95%CI(1.81,2.54)],总有效率高[P0.001,OR=3.26,95%CI (2.5,4.26)],复发率低[P0.001,OR=0.19,95%CI (0.15,0.25)],不良反应发生率低,Begg′s检验表明纳入的RCT研究无发表偏倚(P=0.1020.05)。结论基于现有临床证据,阴道用乳酸菌制剂辅助治疗RVVC疗效优于单纯抗菌药物,可有效改善临床症状,减少疾病发作次数,安全性高,不良反应发生率低,值得临床推广使用。     

人类致病真菌的光谱学检测及鉴定

<正>目前人类致病真菌常规的临床检验方法是直接镜检和真菌分离培养鉴定。直接镜检一般不能确定菌种,培养鉴定费时、费力、阳性率低是其弊端。分子生物学技术受碍于过程繁复、缺乏统一的操作规范和判别标准等,尚未用于临床检验。免疫学的检验方法仅限用于少数致病真菌,且特异性抗体制备不易、昂贵。飞行时间质谱（MALDI-TOF）分析所需设备昂贵,还需分离获得菌株。故人们在不断寻求准确、快速、廉价、高通量、易于标准化的新技术和新方法。     

MALDI-TOF MS技术在临床病原真菌鉴定中的应用进展

基质辅助激光解吸电离飞行时间质谱技术（MALDI-TOF-MS）目前是一种快速而可靠的微生物鉴定方法.随着可鉴定真菌谱的完善,MALDI-TOF MS技术已逐步应用于临床常见致病酵母菌、酵母样真菌和丝状菌的鉴定中,本文将就此做一综述.     

培养条件对克柔念珠菌MALDI-TOF MS鉴定结果的影响分析

目的探究不同的培养时间、培养温度、培养基种类及培养气体环境对克柔念珠菌基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF MS)鉴定结果的影响。方法收集中国侵袭性真菌耐药监测网(CHIF-NET)2012-2013年中17所医院克柔念珠菌25株,每株菌平行接种在沙堡弱氯霉素琼脂平板(SDA-C)、血琼脂平板等6种培养基,28℃和35℃2种孵育温度,空气和5%CO_2 2种气体环境中孵育,以真菌rDNA ITS测序为金标准,孵育24 h和48 h后,使用Vitek MALDI-TOF质谱仪(简称Vitek MS)和Bruker Autoflex Speed型MALDI-TOF质谱仪(简称Bruker MS)分别进行菌种鉴定,并对其鉴定正确率进行分析。结果克柔念珠菌孵育24 h和48 h时Vitek MS鉴定正确率均为100%,Bruker MS鉴定正确率分别是98.86%和87.43%。28℃SDA-C上生长的克柔念珠菌种水平鉴定准确率较35℃生长的菌株高。比较各种培养基,孵育48 h时SDA-C上生长的克柔念珠菌Bruker MS鉴定正确率(100%)最高。结论克柔念珠菌接种在SDA-C上,28℃空气培养24 h质谱鉴定效果最佳。常规细菌培养基上生长的克柔念珠菌,Bruker MS鉴定分值≥1.700即可认为菌种鉴定可靠。     

益生菌治疗类风湿关节炎的meta分析和试验序贯分析

目的 系统评价益生菌治疗类风湿关节炎的临床疗效。方法 检索中国生物医学数据库、中国知网、万方、维普、Embase、PubMed、the Cochrane Library、Web of Science数据库，时间均从建库至2021年7月，对公开发表的益生菌治疗类风湿关节炎的随机对照试验进行meta分析和试验序贯分析。结果 共纳入6项研究，总样本量242例。Meta分析显示，益生菌治疗类风湿关节炎能显著降低CRP[MD=-2.26,95%CI=(-4.30,-0.23),P=0.03]和TNF-α[MD=-1.78,95%CI=(-2.73,0.83),P<0.01]水平，提高IL-10[MD=3.80, 95%CI=(0.40, 7.19), P=0.03]水平，而ESR、 IL-1β、 IL-6、 MDA、 TAC、 DAS28、 TJC、SJC、HAQ和ACR20等水平均与安慰剂相当。敏感性分析显示结果较为稳健，试验序贯分析显示CRP的获益具有结论性，Egger检验显示不存在发表偏倚（P=0.17）。结论 益生菌能够有效降低CRP和TNF-α水平，提高IL-10水平，具有治疗类...     

采用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF)鉴定曲霉属Flavi组和Fumigati组的一些种（英文）

一些曲霉是主要的食物腐败菌及人的病原体,特别是免疫系统受损的病人可能导致严重的感染。本研究评价了利用基质辅助激光解吸电离飞行时间质谱对一些临床和环境中重要的Flavi组和Fumigati组曲霉,进行鉴定的可能性,并将结果与形态学及测序结果(ITS区和部分-tubulin和钙调蛋白基因)进行比较分析。通过曲霉中34个Flavi组菌株和30个Fumigati组菌株的质谱分析,来建立基质辅助激光解吸电离飞行时间质谱的数据库。对光谱数据进行聚类分析表明Fumigati组的基质辅助激光解吸电离飞行时间质谱的结果与系统发育结果完全一致;Flavi组的基质辅助激光解吸电离飞行时间质谱方法将A.flavus,A.oryzae,A.sojae和A.parasiticus分开的效果比测序方法好。随后,再选取用于验证数据库的50个菌株中49个(98%)菌株用质谱数据得到正确鉴定。对于分离本研究中曲霉的隐形种,基质辅助激光解吸电离飞行时间质谱方法优于测序方法。这种方法可以用于曲霉临床实验室鉴定的标准方法,因为临床需要快速和稳定的鉴定方法,这对于适当的治疗方案的选择很重要,此方法同样适于环境研究工作。     

MALDI-TOF MS对临床侵袭性丝状真菌的快速鉴定技术的研究

目的评价基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术在临床常见丝状真菌鉴定中的应用价值。方法收集2017年4月—2019年7月本院各种类型临床标本中分离培养的65株丝状真菌,以及某三甲医院惠赠的26株丝状真菌。这91株丝状真菌通过传统形态学鉴定、DNA测序及MALDI-TOF MS鉴定方法后,以DNA测序结果为标准,与另外2种方法的鉴定结果进行统计学比较,评价MALDI-TOF MS在临床丝状真菌鉴定上的应用。结果在28℃和35℃条件下培养的菌株经质谱仪鉴定后,χ~2检验显示P0.05,提示培养温度差异对结果的影响无统计学意义;以DNA测序结果为金标准,传统形态学方法的正确鉴定率为81.3%(74/91),而基于MALDI-TOF MS技术的正确鉴定率可达97.8%(89/91);研究中,使用"甲酸乙腈提取法"提取蛋白后,丝状真菌质谱鉴定的正确率为90.1%(82/91),而"磁珠研磨法"提取蛋白后的质谱鉴定率则为97.8%(89/91)。结论临床常规工作中使用质谱仪鉴定常见丝状真菌时,培养温度不会影响质谱鉴定结果;改良"磁珠研磨法"提取蛋白后质谱仪鉴定的正确率优于质谱仪推荐"甲酸乙腈提取法";MALDI-TOF MS技术在常见丝状真菌的鉴定上,相比于传统形态学鉴定,该技术更快速、客观、高效及准确。     

ITS序列分析与MALDI-TOF MS质谱技术在丝状真菌鉴定中的应用

丝状真菌常用的鉴定方法为形态方法和基因鉴定方法,前者限于检验人员的知识和技能,后者操作繁琐,费用略昂贵,不适合常规开展。因此,寻找丝状真菌快速鉴定方法势在必行。本文采用VITEK MALDI-TOF MS（基质辅助激光解析电离时间飞行质谱）IVD数据库（3.0版本）对临床分离的254株丝状真菌进行鉴定,并以ITS（internal transcribed spacer 内转录间隔区）序列分析为标准,验证MALDI-TOF MS质谱技术鉴定丝状真菌的准确性。结果表明MALDI-TOF MS质谱技术可以对大部分丝状真菌实现快速、准确的鉴定,其中对毛癣菌属（100%）、毛孢子菌属（100%）、毛霉菌属（100%）、曲霉菌属（96.5%）准确率很高,对犬小孢子菌（75%）、镰刀菌属（50%）、新月弯孢霉（46.2%）准确率较低,对丝状真菌鉴定的总体准确率为86.36%,与ITS测序分析符合率为83.97%。     

Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology

Until recently, microbial identification in clinical diagnostic laboratories has mainly relied on conventional phenotypic and gene sequencing identification techniques. The development of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) devices has revolutionized the routine identification of microorganisms in clinical microbiology laboratories by introducing an easy, rapid, high throughput, low-cost, and efficient identification technique. This technology has been adapted to the constraint of clinical diagnostic laboratories and has the potential to replace and/or complement conventional identification techniques for both bacterial and fungal strains. Using standardized procedures, the resolution of MALDI-TOF MS allows accurate identification at the species level of most Gram-positive and Gram-negative bacterial strains with the exception of a few difficult strains that require more attention and further development of the method. Similarly, the routine identification by MALDI-TOF MS of yeast isolates is reliable and much quicker than conventional techniques. Recent studies have shown that MALDI-TOF MS has also the potential to accurately identify filamentous fungi and dermatophytes, providing that specific standardized procedures are established for these microorganisms. Moreover, MALDI-TOF MS has been used successfully for microbial typing and identification at the subspecies level, demonstrating that this technology is a potential efficient tool for epidemiological studies and for taxonomical classification.     

Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry systems for the identification of clinical filamentous fungi

Infections caused by filamentous fungi have become a health concern, and require rapid and accurate identification in order for effective treatment of the pathogens. To compare the performance of two MALDI-TOF MS systems (Bruker Microflex LT and Xiamen Microtyper) in the identification of filamentous fungal species. A total of 374 clinical filamentous fungal isolates sequentially collected in the Clinical Laboratory at the Beijing Tongren Hospital between January 2014 and December 2015 were identified by traditional phenotypic methods, Bruker Microflex LT and Xiamen Microtyper MALDI-TOF MS, respectively. The discrepancy between these methods was resolved by sequencing for definitive identification. Bruker Microflex LT and Xiamen Microtyper had similar correct species ID (98.9 vs. 99.2%), genus ID (99.7 vs. 100%), mis-ID (0.3 vs. 0%) and no ID (0 vs. 0). The rate of correct species identification by both MALDI-TOF MS (98.9 and 99.2%, respectively) was much higher compared with phenotypic approach (91.9%). Both MALDI-TOF MS systems provide accurate identification of clinical filamentous fungi compared with conventional phenotypic method, and have the potential to replace identification for routine identification of these fungi in clinical mycology laboratories. Both systems have similar performance in the identification of clinical filamentous fungi.     

Evaluation of the Vitek-MS™ system in the identification of Candida isolates from bloodstream infections

Background

Matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF-MS) represents a revolution in the identification of microorganisms of clinical interest. Many studies have confirmed the accuracy and fastness of this tool with routine strains.

A New Filter Based Cultivation Approach for Improving Aspergillus Identification using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS)

Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) is widely used in clinical laboratories for routine identification of bacteria and yeasts. However, methodological difficulties are still apparent when applied to filamentous fungi. The liquid cultivation method recommended by Bruker Daltonics GmbH for identification of filamentous fungi by MALDI-TOF MS is labour intensive and time-consuming. In this study, growth of Aspergillus species on different (porous) surfaces was investigated with the aim to develop a more reliable, quicker and less laborious identification method using MALDI-TOF MS. Mycelial growth without sporulation mimicking liquid cultivation and reliable MALDI-TOF MS spectra were obtained when A. fumigatus strains were grown on and in between a polycarbonate membrane filter on Sabouraud dextrose agar. A database of in-house reference spectra was created by growing Aspergillus reference strains (mainly focusing on sections Fumigati and Flavi) under these selected conditions. A test set of 50 molecularly identified strains grown under different conditions was used to select the best growth condition for identification and to perform an initial validation of the in-house database. Based on these results, the cultivation method on top of a polycarbonate filter proved to be most successful for species identification. This method was therefore selected for the identification of two sets of clinical isolates that mainly consisted of Aspergilli (100 strains originating from Indonesia, 70 isolates from Qatar). The results showed that this cultivation method is reliable for identification of clinically relevant Aspergillus species, with 67% and 76% correct identification of strains from Indonesia and Qatar, respectively. In conclusion, cultivation of Aspergilli on top of a polycarbonate filter showed improved results compared to the liquid cultivation protocol recommended by Bruker in terms of percentage of correct identification, ease of MSP creation, time consumption, cost and labour intensity. This method can be reliably applied for identification of clinically important Aspergilli and has potential for identification of other filamentous fungi.

     

Comparison of the Accuracy of Two Conventional Phenotypic Methods and Two MALDI-TOF MS Systems with That of DNA Sequencing Analysis for Correctly Identifying Clinically Encountered Yeasts

We assessed the accuracy of species-level identification of two commercially available matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) systems (Bruker Biotyper and Vitek MS) and two conventional phenotypic methods (Phoenix 100 YBC and Vitek 2 Yeast ID) with that of rDNA gene sequencing analysis among 200 clinical isolates of commonly encountered yeasts. The correct identification rates of the 200 yeast isolates to species or complex (Candida parapsilosis complex, C. guilliermondii complex and C. rugosa complex) levels by the Bruker Biotyper, Vitek MS (using in vitro devices [IVD] database), Phoenix 100 YBC and Vitek 2 Yeast ID (Sabouraud''s dextrose agar) systems were 92.5%, 79.5%, 89%, and 74%, respectively. An additional 72 isolates of C. parapsilosis complex and 18 from the above 200 isolates (30 in each of C. parapsilosis, C. metapsilosis, and C. orthopsilosis) were also evaluated separately. Bruker Biotyper system could accurately identify all C. parapsilosis complex to species level. Using Vitek 2 MS (IVD) system, all C. parapsilosis but none of C. metapsilosis, or C. orthopsilosis could be accurately identified. Among the 89 yeasts misidentified by the Vitek 2 MS (IVD) system, 39 (43.8%), including 27 C. orthopsilosis isolates, could be correctly identified Using the Vitek MS Plus SARAMIS database for research use only. This resulted in an increase in the rate of correct identification of all yeast isolates (87.5%) by Vitek 2 MS. The two species in C. guilliermondii complex (C. guilliermondii and C. fermentati) isolates were correctly identified by cluster analysis of spectra generated by the Bruker Biotyper system. Based on the results obtained in the current study, MALDI-TOF MS systems present a promising alternative for the routine identification of yeast species, including clinically commonly and rarely encountered yeast species and several species belonging to C. parapsilosis complex, C. guilliermondii complex, and C. rugosa complex.     

A Rapid MALDI-TOF MS Identification Database at Genospecies Level for Clinical and Environmental Aeromonas Strains

The genus Aeromonas has undergone a number of taxonomic and nomenclature revisions over the past 20 years, and new (sub)species and biogroups are continuously described. Standard identification methods such as biochemical characterization have deficiencies and do not allow clarification of the taxonomic position. This report describes the development of a matrix-assisted laser desorption/ionisation–time of flight mass spectrometry (MALDI-TOF MS) identification database for a rapid identification of clinical and environmental Aeromonas isolates.     

MALDI-TOF MS and Filamentous Fungal Identification: A Success Story?

Purpose of Review

The diagnosis of invasive fungal disease remains challenging in the clinical laboratory. In this paper, the use of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of filamentous fungi as well as its application for antifungal resistance testing and strain typing is evaluated.

Recent Findings

Most studies report very high accuracy for the identification of filamentous fungi by MALDI-TOF MS. Its cost effectiveness, short analysis time, and low error rate and the fact that it can also discriminate between closely related and cryptic species make it appropriate for implementation in the clinical routine. Two drawbacks remain in the availability of extended reference spectra databases and the fact that this technique can only be applied on isolates.

Summary

More work on (simultaneous) antifungal susceptibility testing and strain typing is needed. The application of MALDI-TOF MS directly on clinical specimens would further improve the diagnosis of invasive fungal disease and improve its successful management.

     

Accurate Identification of Common Pathogenic Nocardia Species: Evaluation of a Multilocus Sequence Analysis Platform and Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Species identification of Nocardia is not straightforward due to rapidly evolving taxonomy, insufficient discriminatory power of conventional phenotypic methods and also of single gene locus analysis including 16S rRNA gene sequencing. Here we evaluated the ability of a 5-locus (16S rRNA, gyrB, secA1, hsp65 and rpoB) multilocus sequence analysis (MLSA) approach as well as that of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) in comparison with sequencing of the 5’-end 606 bp partial 16S rRNA gene to provide identification of 25 clinical isolates of Nocardia. The 5’-end 606 bp 16S rRNA gene sequencing successfully assigned 24 of 25 (96%) clinical isolates to species level, namely Nocardia cyriacigeorgica (n = 12, 48%), N. farcinica (n = 9, 36%), N. abscessus (n = 2, 8%) and N. otitidiscaviarum (n = 1, 4%). MLSA showed concordance with 16S rRNA gene sequencing results for the same 24 isolates. However, MLSA was able to identify the remaining isolate as N. wallacei, and clustered N. cyriacigeorgica into three subgroups. None of the clinical isolates were correctly identified to the species level by MALDI-TOF MS analysis using the manufacturer-provided database. A small “in-house” spectral database was established incorporating spectra of five clinical isolates representing the five species identified in this study. After complementation with the “in-house” database, of the remaining 20 isolates, 19 (95%) were correctly identified to species level (score ≥ 2.00) and one (an N. abscessus strain) to genus level (score ≥ 1.70 and < 2.00). In summary, MLSA showed superior discriminatory power compared with the 5’-end 606 bp partial 16S rRNA gene sequencing for species identification of Nocardia. MALDI-TOF MS can provide rapid and accurate identification but is reliant on a robust mass spectra database.