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[Comparison of Cytomegalovirus Polymerase Chain Reaction Test Results Obtained with Fully Automated Systems: Roche Cobas 6800 and Qiagen NeuMoDx Experience]. [巨细胞病毒聚合酶链反应检测结果与全自动系统检测结果的比较:罗氏 Cobas 6800 和 Qiagen NeuMoDx 的经验]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20249727
Tuğba Bozdemir, Aylin Erman Daloğlu, Dilek Çolak, Nazlı Gürkan, Gözde Öngüt, Derya Mutlu
<p><p>Viral load monitoring is important in identifying patients at risk of developing cytomegalovirus (CMV) related complications after transplantation and for this purpose, quantitative real-time polymerase chain reaction (Rt-qPCR) tests are most commonly used. The main problem in CMV DNA Rt-qPCR tests that make quantitative measurements is that there are significant differences in measurements performed with different kits in different laboratories. Comparability of viral load measurements between laboratories has increased with the introduction of quantitative PCR tests calibrated with the CMV International Quantitation Standard (IQS) developed by the World Health Organization (WHO). However, quantitative agreement between measurements made with different kits has still not been fully achieved. In this study, it was aimed to investigate the quantitative compatibility between measurements made with Cobas 6800 (Roche Diagnostics, Mannheim, Germany) and NeuMoDx (Qiagen, Ann Arbor, USA) CMV DNA Rt-qPCR tests, which are fully automated new generation systems calibrated with the WHO CMV IQS. The results of 214 plasma samples, which were studied simultaneously with Cobas 6800 CMV Rt-qPCR and NeuMoDx CMV Rt-qPCR tests were analyzed. In the tests, the extraction, amplification and detection stages were carried out fully automatically. CMV DNA was detected in 144 (67.28%) samples in both tests and was not detected in 53 (24.76%) samples. Incompatible results were obtained in a total of 17 (7.94%) samples. Good agreement was found between the qualitative results of both tests (kappa= 0.80, p< 0.001). When the quantitative results (n= 129) obtained in the dynamic measurement range of both tests were examined, the median viral load values measured by Cobas 6800 CMV Rt-qPCR and NeuMoDx CMV Rt-qPCR tests were 513 IU/mL (range= 35-37000) and 741 IU/mL (range= 68-48978), respectively. According to the correlation analysis, a very strong correlation was found between the results of both tests (r= 0.94, p< 0.001). According to Bland-Altman analysis; the average difference between the results of the NeuMoDx CMV Rt-qPCR test and the Cobas 6800 CMV Rt-qPCR test was found to be -0.14 log10 [standard deviation (SD)= 0.23] IU/mL and it was determined that the Cobas 6800 CMV Rt-qPCR test had lower measurements than the NeuMoDx CMV Rt-qPCR test. In 120 of 129 samples (93%) whose results were within the dynamic measurement range of both tests, the measurement difference was within ± 0.5 log10 IU/mL and in 9 (7%), it was detected as more than ± 0.5 log10 (median 0.54 log10 IU/ml; range= 0.51-0.81). No measurement difference of more than ± 1.0 log10 was detected in any sample. In this study, quantitative agreement was found in the measurements made in plasma samples with the fully automated Cobas 6800 CMV Rt-qPCR and NeuMoDx CMV Rt-qPCR tests calibrated with the CMV IQS. To the best of our knowledge, a study comparing viral load measurements made with Cobas 6800 and NeuMo
病毒载量监测对于确定移植后有可能出现巨细胞病毒(CMV)相关并发症的患者非常重要,为此,最常用的是定量实时聚合酶链反应(Rt-qPCR)检测。CMV DNA 实时聚合酶链反应(Rt-qPCR)定量检测的主要问题是,不同实验室使用不同试剂盒进行的检测结果存在显著差异。随着根据世界卫生组织(WHO)制定的 CMV 国际定量标准(IQS)校准的定量 PCR 检测的推出,实验室间病毒载量测量的可比性得到了提高。然而,使用不同试剂盒进行的测量仍未完全达到定量一致。本研究旨在调查 Cobas 6800(罗氏诊断公司,德国曼海姆)和 NeuMoDx(Qiagen,美国安阿伯)CMV DNA Rt-qPCR 检测试剂盒测量结果之间的定量兼容性。对 214 份血浆样本的结果进行了分析,这些样本同时接受了 Cobas 6800 CMV Rt-qPCR 和 NeuMoDx CMV Rt-qPCR 检测。在这些检测中,提取、扩增和检测阶段都是全自动进行的。在这两种检测方法中,有 144 个样本(67.28%)检测到了 CMV DNA,有 53 个样本(24.76%)未检测到 CMV DNA。共有 17 份(7.94%)样本的检测结果不一致。两种检测方法的定性结果具有良好的一致性(kappa= 0.80,p< 0.001)。如果对两种检测方法在动态测量范围内获得的定量结果(n= 129)进行检查,Cobas 6800 CMV Rt-qPCR 和 NeuMoDx CMV Rt-qPCR 检测方法测得的病毒载量中位值分别为 513 IU/mL(范围= 35-37000)和 741 IU/mL(范围= 68-48978)。根据相关性分析,两种检测结果之间存在很强的相关性(r= 0.94,p< 0.001)。根据Bland-Altman分析,NeuMoDx CMV Rt-qPCR检测结果与Cobas 6800 CMV Rt-qPCR检测结果的平均差异为-0.14 log10 [标准差(SD)= 0.23] IU/mL,并确定Cobas 6800 CMV Rt-qPCR检测结果低于NeuMoDx CMV Rt-qPCR检测结果。在 129 份样本中,有 120 份样本(93%)的结果在两种检测方法的动态测量范围内,其测量差异在 ± 0.5 log10 IU/ml 以内,有 9 份样本(7%)的测量差异超过 ± 0.5 log10(中位数 0.54 log10 IU/ml;范围= 0.51-0.81)。在所有样本中均未发现测量值差异超过 ± 1.0 log10 的情况。在本研究中,全自动 Cobas 6800 CMV Rt-qPCR 和 NeuMoDx CMV Rt-qPCR 检测仪与 CMV IQS 校准血浆样本的定量测量结果一致。据我们所知,在检测 CMV DNA 时比较 Cobas 6800 和 NeuMoDx 全自动系统的病毒载量测量结果的研究还没有进行过,本研究是这方面的第一项研究。
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引用次数: 0
[Evaluation of the Bio-Speedy Meningitis/Encephalitis Panel in the Diagnosis of Central Nervous System Infections]. [评估 Bio-Speedy 脑膜炎/脑炎样本在诊断中枢神经系统感染中的作用]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.202497189
Osman Merdan, İmran Sağlık, Fatma Dilşad Aksoy, Uğur Önal, Cüneyt Özakın, Mustafa Kemal Hacımustafaoğlu, Solmaz Çelebi, Harun Ağca, Mehmet Tekinsoy, Beyza Ener
<p><p>Infections of the central nervous system (CNS) can lead to severe outcomes if not accurately diagnosed and treated. The broad spectrum of pathogens involved in CNS infections can make diagnosis challenging. Polymerase chain reaction (PCR) -based multiplex molecular diagnostic panels can rapidly and simultaneously detect multiple neuropathogens in cerebrospinal fluid (CSF). This study was aimed to assess the Bio-Speedy Meningitis/Encephalitis RT-PCR MX-17 panel (Bioeksen, İstanbul, Türkiye), a novel multiplex PCR test, in diagnosing CNS infections. The panel can detect a range of pathogens, including Escherichia coli K1, Haemophilus influenzae, Listeria monocytogenes, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus agalactiae, enterovirus (EV), herpes simplex virus (HSV) 1 and 2, HHV-6, HHV-7, HHV-8, human parechovirus (HPeV), varicella zoster virus (VZV), cytomegalovirus (CMV) and Cryptococcus gatti/neoformans in CSF samples. This retrospective study included 128 CSF samples from 128 patients sent to Bursa Uludağ University Health Application and Research Center Microbiology Laboratory between June 2022 and July 2023 to search for CNS infectious agents. Patient clinical, radiological and laboratory data were collected from the Hospital Information Record System (HIRS). Bacterial pathogens were identified through culture, while viral pathogens were detected in CSF samples using the Fast Track Diagnostics (FTD) multiplex RT-PCR panel (Fast Track Diagnostics Ltd., Luxembourg) for HSV-1, HSV-2, VZV, EV, mumps virus and HPeV. The stored CSF samples were then tested using the BioSpeedy panel and the results were compared with those of the culture and the FTD panel. Pathogens that were detected were considered positive if they were consistent with the patient's symptoms and CSF characteristics according to infectious disease and pediatric infectious disease specialists. Pathogens detected but not supported by the patient's symptoms and CSF characteristics were classified as uncertain clinical relevance (UCR). Out of the 128 patients tested for CNS infectious agents, 44 (34.4%) were diagnosed with a CNS infection. The overall pathogen detection rate with all methods was 43.2% (19/44). The Bio-Speedy panel identified pathogens in 29.5% (13/44) of the patients, followed by the FTD panel (20.5%, 9/44) and culture (9.1%, 4/44). Four bacteria were identified with culture, three of which were also detected by the Bio-Speedy panel. Additionally, six bacteria were identified with Bio-Speedy panel, that were not identified by culture. The FTD panel identified nine viruses, four of which were also identified by Bio-Speedy. In total, the Bio-Speedy panel detected 13 of the 19 positive pathogens (nine bacteria and four viruses: [S.pneumoniae (n= 3), VZV (n= 3), N.meningitidis (n= 2), H.influenzae (n= 2), L.monocytogenes (n= 1), E.coli (n= 1) ve EV (n= 1)]. However, the Bio-Speedy panel identified 15 pathogens [S.pneumoniae (n= 1), E.coli (n= 1)
中枢神经系统(CNS)感染如果得不到准确诊断和治疗,可能会导致严重后果。中枢神经系统感染的病原体种类繁多,这给诊断带来了挑战。基于聚合酶链式反应(PCR)的多重分子诊断试剂盒可同时快速检测脑脊液(CSF)中的多种神经病原体。本研究旨在评估 Bio-Speedy 脑膜炎/脑炎 RT-PCR MX-17 检验板(Bioeksen,伊斯坦布尔,图尔基耶)在诊断中枢神经系统感染方面的作用。该试剂盒可检测一系列病原体,包括大肠杆菌 K1、流感嗜血杆菌、单核细胞增生李斯特菌、脑膜炎奈瑟菌、肺炎链球菌、无乳链球菌、肠道病毒 (EV)、疱疹病毒 (EV)、疱疹病毒 (EV)、肠道病毒 (EV)、单纯疱疹病毒 (HSV) 1 和 2、HHV-6、HHV-7、HHV-8、人类帕累托病毒 (HPeV)、水痘带状疱疹病毒 (VZV)、巨细胞病毒 (CMV) 和加蒂隐球菌/新变形隐球菌。这项回顾性研究纳入了 2022 年 6 月至 2023 年 7 月间送往布尔萨乌鲁达大学健康应用与研究中心微生物实验室的 128 例患者的 CSF 样本,以寻找中枢神经系统感染病原体。患者的临床、放射和实验室数据均来自医院信息记录系统(HIRS)。细菌病原体通过培养进行鉴定,病毒病原体则通过快速诊断公司(FTD)的多重 RT-PCR 小组(Fast Track Diagnostics Ltd., Luxembourg)对 CSF 样本中的 HSV-1、HSV-2、VZV、EV、腮腺炎病毒和 HPeV 进行检测。然后使用 BioSpeedy 检测板对储存的 CSF 样品进行检测,并将结果与培养和 FTD 检测板的结果进行比较。根据传染病专家和儿科传染病专家的意见,如果检测出的病原体与患者的症状和 CSF 特征相符,则认为是阳性病原体。检测到的病原体如果与患者症状和 CSF 特征不符,则被归类为临床相关性不确定(UCR)。在接受中枢神经系统感染病原体检测的 128 名患者中,有 44 人(34.4%)被确诊为中枢神经系统感染。所有方法的病原体总检出率为 43.2%(19/44)。Bio-Speedy小组在29.5%(13/44)的患者中发现了病原体,其次是FTD小组(20.5%,9/44)和培养(9.1%,4/44)。通过培养确定了四种细菌,其中三种也被 Bio-Speedy 小组检测到。此外,Bio-Speedy 鉴定小组还鉴定出 6 种细菌,但培养方法未鉴定出这些细菌。FTD 检测板检测出 9 种病毒,其中 4 种也被 Bio-Speedy 检测板检测出来。在 19 种阳性病原体中,Bio-Speedy 小组共检测出 13 种(9 种细菌和 4 种病毒):[肺炎双球菌(3 个)、VZV(3 个)、脑膜炎双球菌(2 个)、流感嗜血杆菌(2 个)、单核细胞增多性乳酸杆菌(1 个)、大肠杆菌(1 个)和 EV(1 个)]。然而,Bio-Speedy 鉴定小组确定了 15 种病原体[肺炎链球菌(n=1)、大肠杆菌(n=1)、卡地/新变形杆菌(n=1)、巨细胞病毒(n=8)、HHV-6(n=3)和 HHV-7 (n=1)]被认为是 UCR。在确诊的中枢神经系统感染患者中,Bio-Speedy 鉴定出致病病原体的比例最高(29.5%)。尽管如此,仍应根据患者的特征来解释检测结果,以确保对患者进行适当的管理。使用多种方法和多重检验可提高中枢神经系统感染的诊断准确性。
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引用次数: 0
[Seroprevalence and Risk Factors of Hepatitis B, C and D in Adults in Trabzon, Türkiye]. [土耳其特拉布宗成人乙型、丙型和丁型肝炎血清阳性率和风险因素]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20249762
Cevriye Ceyda Kolaylı, Murat Topbaş, Esra Özkaya, İftihar Köksal, Nazım Ercüment Beyhun, Neşe Kaklıkkaya, Gamze Çan, Mustafa Yılmaz, Köksal Hamzaoğlu, Esin Sayın, Serdar Karakullukçu

Viral hepatitis are infections that can cause liver damage, become chronic, lead to cirrhosis, hepatocellular carcinoma and ultimately result in death due to their ability to spread in the community through blood and infected body fluids. The aim of this study was to determine the seroprevalence of hepatitis B (HBV), hepatitis C (HCV), and hepatitis D (HDV) transmitted through blood among individuals living in Trabzon province and to examine the factors potentially associated with seroprevalence. This cross-sectional study was conducted in Trabzon province, located in the northeast of Türkiye, including a total of 10 districts, including the central district. Since seroprevalence was calculated for HBV, HCV, and HDV in the study, the sample size was separately calculated for each, and the calculated maximum sample size of 1116 was accepted as the minimum sample size for the study. The study was completed with 1502 participants. Serological tests for HBV included HBsAg, anti-HBs, and anti-HBc IgG; for HCV, anti-HCV; and for HDV, anti-HDV were analysed. Data were evaluated for HBV risk factors using univariate analyses with Chi-square test and for multiple analyses using enter model logistic regression analysis. The mean age of the participants was 45.7 ± 16.6 years, with 767 (51.1%) being female. The prevalence of HBV seropositivity, indicating vaccination, was 23.0%, while the seroprevalence of HBV among unvaccinated adults was 27.4%. HBsAg positivity was 5.1%, and isolated anti-HBc IgG positivity was 4.2%. The proportion of individuals with HBsAg in the gray zone was 0.5%, while the positivity rates for anti-HBs and anti-HBc IgG (indicating past infection) were 17.6%. The prevalence of anti-HCV was six per thousand, while anti-HDV was not detected in the analyses. HBsAg positivity and co-infection with HCV were found in one person, and among the nine individuals positive for anti-HCV, isolated anti-HBc IgG positivity was detected in three. Increasing age, presence of a person with jaundice in the family, presence of diabetes mellitus, alcohol use and cupping therapy were identified as risk factors for HBV in the logistic regression analysis. Risk factors for HCV in univariate analyses were being over 40 years old, presence of hepatic steatosis and receiving dialysis treatment. The results of the study indicate that despite being included in our vaccination schedule and the administration of vaccines to high-risk adults, HBV still requires intensive attention as a public health problem. HCV, lacking a vaccine has been evaluated as an infectious agent that needs to be taken into consideration due to its potential risks and requires the complete implementation of individual and social precautions.

病毒性肝炎是一种可导致肝损伤、慢性化、导致肝硬化、肝细胞癌并最终导致死亡的感染,因为它们能够通过血液和受感染的体液在社区中传播。本研究旨在确定特拉布宗省居民中通过血液传播的乙型肝炎(HBV)、丙型肝炎(HCV)和丁型肝炎(HDV)的血清流行率,并研究与血清流行率潜在相关的因素。这项横断面研究在特拉布宗省进行,该省位于土耳其东北部,包括中部地区在内共有 10 个县。由于研究计算的是 HBV、HCV 和 HDV 的血清流行率,因此分别计算了每种病毒的样本量,并将计算出的最大样本量 1116 人作为研究的最小样本量。该研究共有 1502 名参与者。HBV 血清学检测包括 HBsAg、抗-HBs 和抗-HBc IgG;HCV 血清学检测包括抗-HCV;HDV 血清学检测包括抗-HDV。对数据进行了HBV风险因素评估,单变量分析采用Chi-square检验,多重分析采用enter模型逻辑回归分析。参与者的平均年龄为 45.7 ± 16.6 岁,其中 767 人(51.1%)为女性。接种疫苗后,HBV 血清阳性率为 23.0%,而未接种疫苗的成年人的 HBV 血清阳性率为 27.4%。HBsAg 阳性率为 5.1%,分离抗 HBc IgG 阳性率为 4.2%。灰色区域的 HBsAg 阳性率为 0.5%,抗-HBs 和抗-HBc IgG 阳性率为 17.6%(表明既往感染)。抗-HCV 感染率为千分之六,而抗-HDV 在分析中未检测到。有一人发现 HBsAg 阳性并同时感染了 HCV,而在抗-HCV 阳性的九人中,有三人检测到单独的抗-HBc IgG 阳性。在逻辑回归分析中,年龄增加、家族中有黄疸患者、糖尿病患者、饮酒和拔罐治疗被确定为 HBV 的风险因素。在单变量分析中,40 岁以上、肝脏脂肪变性和接受透析治疗是感染 HCV 的风险因素。研究结果表明,尽管 HBV 已被纳入我们的疫苗接种计划,并对高危成人进行了疫苗接种,但作为一个公共卫生问题,HBV 仍需要引起高度重视。丙型肝炎病毒(HCV)没有疫苗,但由于其潜在风险,已被评估为一种需要考虑的传染源,需要全面落实个人和社会预防措施。
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引用次数: 0
[Effect of Quorum Sensing Systems on Biofilm Formation and Virulence in Salmonella]. [法定人数感应系统对沙门氏菌生物膜形成和毒力的影响]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20240038
Ebru Öztaş Gülmüş, Nefise Akçelik, Caner Özdemir, Mustafa Akçelik

In recent years, as the paradigm of communication between cells has been clarified, the ability of bacteria to change their gene expression patterns in response to various extracellular signals has attracted great interest. In particular, intracellular and intercellular communication between bacterial populations, called quorum sensing (QS), is essential for coordinating physiological and genetic activities. QS studies are critical, particularly in elucidating the regulatory mechanisms of infectious processes in food-borne pathogens. Elucidating the QS mechanisms in Salmonella is effective in silencing the virulence factors in the fight against this bacterium. The aims of this study were; to create luxS gene mutants that play a vital role in the QS activity of Salmonella and to determine the effect of this mutation on the expression of virulence genes in the bacteria and to determine the impact of synthetic N-hexanoyl-homoserine lactone (C6HSL) on biofilm formation and AI-2 signaling pathway of Salmonella wild strain and luxS gene mutants. luxS gene mutants were constructed by recombining the gene region with the chloramphenicol gene cassette based on homologous region recombination. In the luxS mutants obtained in this way, the expression of eight different virulence genes (hilA, invA, inv, glgC, fimF, fliF, lpfA, gyrA), which have essential roles in Salmonella pathogenicity, was determined by quantitative real-time reverse transcriptase polymerase chain reaction (rRT-qPCR) method and compared with natural strains. As a result of these studies, it was determined that the expression of each gene examined was significantly reduced in luxS mutant strains. The relative AI-2 activities of Salmonella strains were analyzed depending on time. It was determined that the highest activity occurred at the fourth hour and the AI-2 activities of luxS mutants were reduced compared to the wild strain. Finally, it was determined that C6HSL increased the biofilm activity of Salmonella Typhimurium DMC4, SL1344 wild strains, and mutants, mainly at the 72nd hour. In conclusion, our results proved that C6HSL stimulated QS communication in all strains and increased biofilm of Salmonella formation and autoinducer activity. This situation determines that Salmonella responds to external signals by using QS systems. In addition, this research contributed to provide additional information on interspecies communication mechanisms to develop strategies to prevent biofilm formation of this pathogen.

近年来,随着细胞间通信模式的明确,细菌根据各种细胞外信号改变基因表达模式的能力引起了人们的极大兴趣。其中,细菌种群之间的胞内和胞间通信(称为法定量感应(QS))对于协调生理和遗传活动至关重要。QS 研究至关重要,尤其是在阐明食源性病原体感染过程的调控机制方面。阐明沙门氏菌的 QS 机制可有效抑制该细菌的毒力因子。本研究的目的是:创建在沙门氏菌QS活性中发挥重要作用的luxS基因突变株,并确定该突变对细菌毒力基因表达的影响,以及确定合成N-己酰-高丝氨酸内酯(C6HSL)对沙门氏菌野生菌株和luxS基因突变株的生物膜形成和AI-2信号通路的影响。luxS基因突变株是根据同源区重组法将该基因区与氯霉素基因盒重组而构建的。通过实时反转录聚合酶链式反应(rRT-qPCR)定量测定了在沙门氏菌致病性中起重要作用的8种不同毒力基因(hilA、invA、inv、glgC、fimF、fliF、lpfA、gyrA)在luxS基因突变株中的表达情况,并与天然菌株进行了比较。研究结果表明,在luxS突变菌株中,每个被检测基因的表达量都显著降低。沙门氏菌菌株的相对 AI-2 活性随时间变化而分析。结果表明,在第四个小时时活性最高,与野生菌株相比,luxS突变体的AI-2活性降低。最后,C6HSL 提高了鼠伤寒沙门氏菌 DMC4、SL1344 野生菌株和突变体的生物膜活性,主要是在第 72 小时。总之,我们的研究结果证明,C6HSL 可刺激所有菌株的 QS 通信,增加沙门氏菌生物膜的形成和自诱导活性。这种情况表明,沙门氏菌利用 QS 系统对外部信号做出反应。此外,这项研究还有助于提供更多关于种间通讯机制的信息,以制定策略防止这种病原体形成生物膜。
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引用次数: 0
[Investigation of Biofilm Formation, Anti-Quorum Sensing Activity and Antimicrobial Resistance in Corynebacterium Species Isolated from Clinical Samples]. [从临床样本中分离出的棒状杆菌的生物膜形成、抗法定人数感应活性和抗菌药耐药性研究]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20249704
Banu Hümeyra Keskin, İdris Şahin, Gözde Kahraman, Pelin Kamuran Duran, Görkem Dülger, Mehmet Akif Durmuş, Ayşe Nur Ceylan, Emel Çalışkan, Şükrü Öksüz
<p><p>An increasing number of different clinical infections caused by Corynebacteria have been reported in the last decade. The aim of this study was to evaluate the antibiotic resistance rates, biofilm formation capacities and to investigate the ''anti-quorum-sensing (anti-QS)'' activities of corynebacteria, which were divided into three groups according to the type of growth in culture (pure, with another pathogenic bacterium and polymicrobial growth). In total 240 Corynebacterium spp. isolates from different clinical specimens sent to the medical microbiology laboratories of Düzce University Faculty of Medicine Hospital and Başakşehir Çam and Sakura City Hospital between June 2021 and June 2022 were classified into three groups: pure, isolated with another pathogen and polymicrobial, according to their growth patterns in culture. Bacteria were identified by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) Biotyper (Bruker, Germany) at an external centre. Antibiotic susceptibilities were determined by disc diffusion method and for vancomycin broth microdilution method was used. Results were interpreted according to EUCAST recommendations. The biofilm-forming properties of the isolates were determined quantitatively. Bioactive components of 17 isolates with strong biofilm formation were extracted and anti-QS activity was determined by agar diffusion method using Chromobacterium violaceum ATCC 12472 strain and then violacein pigment production was measured quantitatively. Of the 240 Corynebacterium spp. isolates, 138 (58%) were pure, 52 (22%) were isolated with another pathogen and 50 (20%) were part of a polymicrobial infection. Of the isolates, 140 were identified as C.striatum, 34 as C.amycolatum and 24 as Corynebacterium afermentans. When the antibiotic resistance rates of the Corynebacterium isolates were analysed according to the groups, the resistance rates to rifampicin and tetracycline antibiotics were found to be statistically significantly lower in the polymicrobial group than in the other groups. The resistance rates to penicillin, clindamycin, ciprofloxacin, moxifloxacin, rifampicin, tetracycline and linezolid were 96.7%, 88.3%, 86.3%, 73.8%, 62.5%, 59.2% and 0.8%, respectively. While all isolates were susceptible to vancomycin, linezolid resistance was detected in two C.afermentans isolates. When the biofilm formation ability was analysed, it was observed that 87 (36.3%) isolates formed biofilm. The biofilm formation rate of the isolates in the polymicrobial growth group was lower than the other two groups. The anti-QS activity of 17 isolates with strong biofilm formation was investigated and none of the Corynebacterium extracts tested were found to have anti-QS activity (inhibition of violacein pigment production without inhibiting bacterial growth) in the QS study with C.violaceum, whereas five isolate extracts had antibacterial activity (inhibition of bacterial growth). Four of the bacte
近十年来,由棒状杆菌引起的各种临床感染的报道越来越多。本研究的目的是评估棒状杆菌的抗生素耐药率、生物膜形成能力,并研究棒状杆菌的 "抗法定量感应(anti-QS)"活性,根据培养物的生长类型(纯生长、与其他致病菌一起生长和多微生物生长)将棒状杆菌分为三组。2021 年 6 月至 2022 年 6 月期间,杜塞大学医学院附属医院和巴沙克谢希尔恰姆和樱花市医院医学微生物实验室从不同的临床标本中分离出 240 株棒状杆菌,根据培养物的生长模式将其分为三组:纯合菌、与其他病原体分离菌和多微生物菌。细菌在外部中心通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)Biotyper(德国布鲁克公司)进行鉴定。抗生素敏感性采用盘扩散法测定,万古霉素采用肉汤微量稀释法测定。结果根据欧盟食品和药品检验委员会(EUCAST)的建议进行解释。对分离物的生物膜形成特性进行了定量测定。提取了 17 个具有较强生物膜形成能力的分离菌株的生物活性成分,并使用 Chromobacterium violaceum ATCC 12472 菌株通过琼脂扩散法测定了抗 QS 活性,然后定量测定了暴力素色素的产生。在 240 株棒状杆菌属分离物中,138 株(58%)是纯种的,52 株(22%)是与其他病原体一起分离的,50 株(20%)是多微生物感染的一部分。在这些分离物中,140 个被鉴定为 C.striatum,34 个被鉴定为 C.amycolatum,24 个被鉴定为 Corynebacterium afermentans。根据不同组别对分离出的棒状杆菌的抗生素耐药率进行分析后发现,多菌感染组对利福平和四环素类抗生素的耐药率在统计学上明显低于其他组别。对青霉素、克林霉素、环丙沙星、莫西沙星、利福平、四环素和利奈唑胺的耐药率分别为 96.7%、88.3%、86.3%、73.8%、62.5%、59.2% 和 0.8%。虽然所有分离株都对万古霉素敏感,但在两个 C.afermentans 分离株中检测到了利奈唑胺耐药性。在分析生物膜形成能力时,发现有 87 个(36.3%)分离物形成了生物膜。多微生物生长组分离物的生物膜形成率低于其他两组。对 17 个生物膜形成较强的分离物的抗 QS 活性进行了调查,在对 C.violaceum 的 QS 研究中,没有发现任何一种被测试的棒状杆菌提取物具有抗 QS 活性(抑制长春花苷色素的产生而不抑制细菌生长),而有 5 种分离物提取物具有抗菌活性(抑制细菌生长)。在具有抗菌活性的细菌提取物中,4 种属于 C.amycolatum,1 种属于 C.afermentans。总之,在对抗生素耐药率和生物膜形成率进行分析时,与其他病原体一起培养的伞菌与纯培养的伞菌表现出相似的特征。因此,我们认为不应忽视与另一种病原体一起生长的棒状杆菌。此外,一些伞菌提取物的抗菌效果表明,应该对微生物群细菌进行更多的质量控制研究。
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引用次数: 0
[Aspergillus flavus/oryzae Arthritis of the Knee Joint: First Case in Türkiye]. [膝关节黄曲霉菌/oryzae 关节炎:土耳其首例]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20249702
Selda Kömeç, İlgin Özden
<p><p>Aspergillus species are common hyphal fungi. In addition to allergies and mycotoxicosis, Aspergillus species can cause various infections known as aspergillosis. Aspergillosis of the respiratory tract, central nervous system, skin and soft tissues is well described. However, musculoskeletal infections due to invasive aspergillosis are not well described. Fungal joint infection due to invasive aspergillosis is a rare form of septic arthritis. In this case report, a patient who admitted to our hospital for liver transplantation and developed knee joint arthritis caused by Aspergillus flavus/Aspergillus oryzae during this process was presented. A 28-year-old male patient with autoimmune hepatitis was admitted to hospital with decompensated liver cirrhosis and encephalopathy. The patient, who was awaiting an emergency liver transplant, developed pain, swelling and limitation of movement in his right knee and appropriate consultations and tests were requested. Three joint fluid cultures taken one day apart and nine days later were positive for fungal growth. Macroscopic examination of the mould growth and microscopic examination with lactophenol cotton blue suggested a species belonging to the A.flavus complex and the isolate was identified as A.flavus/A.oryzae by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF MS) (EXS 2600, Zybio, China). As a result of ITS gene sequencing, the species was determined to be A.oryzae. As cases have been reported where A.flavus and A.oryzae species could not be distinguished by ITS gene sequencing, the pathogen was defined as A.flavus/oryzae. The patient died of liver disease during treatment with amphotericin B. There are few cases of arthritis caused by Aspergillus species in the literature. Aspergillus species found in joint infections are, Aspergillus fumigatus, A.flavus, Aspergillus niger and Aspergillus terreus species complexes, in order of frequency. A.flavus and A.oryzae are closely related. They are difficult to distinguish by conventional methods, MALDI-TOF MS or ITS region sequencing, which is commonly used for genus/species identification in fungi. The number of Aspergillus arthritis cases is low and the identification methods applied to the species reported as causative agents in most studies can identify at the species complex level. In addition, it can be assumed that species not previously reported as causative agents may be encountered as a result of developments in identification methods. In the few publications in the literature where A.flavus complex was reported as the causative agent of joint infections, it seems possible that some of the agents may be A.flavus and some may be A.oryzae, since the agents were identified at the complex level. There are a limited number of cases in the literature where A.oryzae is the causative agent, particularly in the respiratory tract. A PubMed search using the keywords "A.oryzae infections, arthritis, osteomyelitis" did not reveal
曲霉菌是一种常见的半知菌。除了过敏和霉菌中毒外,曲霉菌还可引起各种感染,即曲霉菌病。呼吸道、中枢神经系统、皮肤和软组织的曲霉菌病已被详细描述。然而,由侵袭性曲霉菌病引起的肌肉骨骼感染却没有得到很好的描述。侵袭性曲霉菌病引起的真菌性关节感染是一种罕见的化脓性关节炎。在本病例报告中,我们介绍了一名入院接受肝脏移植手术的患者,他在手术过程中因黄曲霉菌/奥氏曲霉菌引起了膝关节关节炎。一名患有自身免疫性肝炎的 28 岁男性患者因肝硬化失代偿期和脑病入院。患者正在等待紧急肝脏移植手术,他的右膝盖出现疼痛、肿胀和活动受限,因此要求进行适当的会诊和检查。相隔一天和九天后进行的三次关节液培养均显示真菌生长呈阳性。霉菌生长的宏观检查和用乳酚棉蓝进行的显微镜检查表明,该菌株属于黄曲霉菌群,并通过基质辅助激光解吸电离质谱(MALDI-TOF MS)(EXS 2600,Zybio,中国)鉴定为黄曲霉/奥里泽。通过 ITS 基因测序,确定该物种为 A.oryzae。ITS 基因测序无法区分黄曲霉和奥里泽霉的病例也有报道,因此病原体被定义为黄曲霉/奥里泽霉。该患者在接受两性霉素 B 治疗期间死于肝病。在关节感染中发现的曲霉菌依次为烟曲霉、黄曲霉、黑曲霉和土曲霉菌群。黄曲霉和黑曲霉关系密切。传统方法、MALDI-TOF MS 或 ITS 区域测序(通常用于真菌属/种的鉴定)都很难将它们区分开来。曲霉关节炎病例的数量较少,而大多数研究中对报告为致病菌的菌种所采用的鉴定方法可以在菌种复合水平上进行鉴定。此外,我们还可以假设,由于鉴定方法的发展,可能会遇到以前未被报告为致病菌的菌种。在少数报道黄曲霉菌复合体是关节感染病原体的文献中,由于病原体是在复合体水平上鉴定的,因此似乎有些病原体可能是黄曲霉菌,有些可能是oryzae。在文献中,A.oryzae 是致病菌的病例数量有限,尤其是在呼吸道。在PubMed上以 "A.oryzae感染、关节炎、骨髓炎 "为关键词进行搜索,没有发现任何关于A.oryzae引起关节感染的文献。
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引用次数: 0
[Investigation of hsa-miR-1908 and hsa-miR-144 Expression Levels in Crimean-Congo Hemorrhagic Fever Patients]. [克里米亚-刚果出血热患者体内 hsa-miR-1908 和 hsa-miR-144 表达水平的调查]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.202497188
Özlem Aldemir, Aynur Engin, Burcu Bayyurt, Serdal Arslan

Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic viral disease. MicroRNAs (miRNAs), which play an important role in the regulation of gene expression, are involved in many processes essential for cell life such as development, differentiation, survival, apoptosis and aging. If miRNAs fail to fulfill their functions, they cause susceptibility to many diseases, including viral infections or cause the disease to be experienced in different clinical situations, such as severe or mild. In this study, we aimed to determine the expression levels of hsa-miR-144 and hsa-miR-1908 in CCHF patients and to compare the results in different clinical courses of CCHF disease. In this study, expression levels of hsa-miR-144 and hsa-miR-1908 were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR) in blood samples obtained from 60 CCHF patients and 40 healthy individuals. We also investigated the differences in the expression levels of the microRNAs between patients with severe and non-severe disease or between patients who died and survived. Quantitative polymerase chain reaction data were uploaded to the "Data Analysis Center" (Qiagen, Germany) and analyzed using the ΔΔCq (ΔΔCt) method. p value was calculated according to Student's t test for genes in the study groups. The expression level of hsa-miR-144 decreased (fold change= 0.09) and the expression level of hsa-miR-1908 increased 1.44-fold in CCHF patients compared to the control group. The expression of hsa-miR-144 and hsa-miR-1908 increased 2 and 2.36-fold, respectively, in severe patients compared to non-severe patients. The expression levels of hsa-miR-144 and hsa-miR-1908 were 16.3- and 14.3-fold higher, respectively, in fatal cases compared to surviving patients and these results were statistically significant. In addition, the expression level of hsa-miR-144 was significantly decreased in patients with low leukocyte counts and the expression level of hsa-miR-1908 was significantly increased in patients with prolonged prothrombin time (PT). This is the first study in the literature investigating the expression level of hsa-miR-1908 in CCHF patients. In conclusion, the data of this study suggest that hsa-miR-144 and hsa-miR-1908 may be important biomarkers in predicting the prognosis and clinical course of CCHF disease.

克里米亚-刚果出血热(CCHF)是一种蜱传人畜共患病毒性疾病。微小核糖核酸(miRNA)在调控基因表达方面发挥着重要作用,参与细胞生命的许多重要过程,如发育、分化、存活、凋亡和衰老。如果 miRNAs 不能发挥其功能,就会导致许多疾病的易感性,包括病毒感染,或导致疾病在不同的临床情况下发生,如严重或轻微。本研究旨在测定 hsa-miR-144 和 hsa-miR-1908 在 CCHF 患者中的表达水平,并比较其在 CCHF 疾病不同临床过程中的结果。本研究通过定量反转录聚合酶链反应(RT-qPCR)检测了 60 名 CCHF 患者和 40 名健康人血液样本中 hsa-miR-144 和 hsa-miR-1908 的表达水平。我们还研究了重症和非重症患者之间或死亡和存活患者之间 microRNA 表达水平的差异。定量聚合酶链反应数据上传到 "数据分析中心"(Qiagen,德国),并使用ΔΔCq(ΔΔCt)方法进行分析。与对照组相比,CCHF 患者 hsa-miR-144 的表达水平下降(折合变化= 0.09),hsa-miR-1908 的表达水平上升 1.44 倍。与非重度患者相比,重度患者中 hsa-miR-144 和 hsa-miR-1908 的表达分别增加了 2 倍和 2.36 倍。在死亡病例中,hsa-miR-144 和 hsa-miR-1908 的表达水平分别是存活病例的 16.3 倍和 14.3 倍,这些结果具有统计学意义。此外,在白细胞计数低的患者中,hsa-miR-144 的表达水平明显降低,而在凝血酶原时间(PT)延长的患者中,hsa-miR-1908 的表达水平明显升高。这是文献中首次研究 hsa-miR-1908 在 CCHF 患者中的表达水平。总之,本研究的数据表明,hsa-miR-144 和 hsa-miR-1908 可能是预测 CCHF 疾病预后和临床过程的重要生物标志物。
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引用次数: 0
[A New Method for Determination of Rifampicin and Isoniazid Resistance in Mycobacterium tuberculosis complex Isolates: Capillary Tube Method]. [一种测定结核分枝杆菌复合菌株对利福平和异烟肼耐药性的新方法:毛细管法]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20249764
Nazlı Arslan, Ebru Demiray Gürbüz, Ayşe Aydan Özkütük, Nuran Esen

Tuberculosis continues to be an important public health problem worldwide. Culture methods are still considered the gold standard in the diagnosis of tuberculosis and the determination of drug resistance. The most important limitation of these methods is their long turnaround time. Commercial culture systems developed to shorten the duration are emerging as an economic problem, especially for developing countries. Therefore, cheap, fast, easy to apply and objectively evaluable tests are needed. In this study, in addition to culture-based methods for determining RIF and INH resistance in Mycobacterium tuberculosis complex isolates, it was aimed to develop the capillary tube method to accelerate the evaluation process. The study included 27 RIF-resistant, 36 RIF -sensitive, 30 INH-resistant, and 33 INH-sensitive isolates obtained from the mycobacteriology laboratory culture collection, for which susceptibility testing to firstline drugs were previously performed using the BACTEC MGIT 960 system (BD, USA) and were stored. H37Rv standard strain and an external quality control strain (IDT3) with known RIF and INH resistance were used as quality control isolates in the study. As a new testing method, the capillary tube method for detecting rifampicin and isoniazid resistance was compared to the standard BACTEC MGIT 960 system. In the determination of RIF and INH resistance, the sensitivity of the capillary tube method compared to the reference method was determined as 85% and 80%, respectively; however, the specificity values (25% and 45.5%, respectively) for both drugs were found to be low in the studies. The time to detect resistance with the capillary tube method varied between 4-9 days. Capillary tube method, which was developed especially for the rapid identification and treatment of multidrug-resistant isolates, is promising in that it detects resistant strains in a short time with a relatively high sensitivity, although its specificity is very low. It is thought that it would be beneficial to continue the study with a larger number of samples and even improve the method with studies conducted directly from clinical samples.

结核病仍然是全球重要的公共卫生问题。培养方法仍被认为是诊断结核病和确定耐药性的黄金标准。这些方法最重要的局限是周转时间长。为缩短周转时间而开发的商业培养系统正在成为一个经济问题,尤其是对发展中国家而言。因此,需要廉价、快速、易于应用且可客观评价的检测方法。在本研究中,除了用培养法测定复合结核分枝杆菌对 RIF 和 INH 的耐药性外,还旨在开发毛细管法,以加快评估过程。研究对象包括从结核分枝杆菌实验室培养物收集中获得的 27 株 RIF 耐药株、36 株 RIF 敏感株、30 株 INH 耐药株和 33 株 INH 敏感株分离物,这些分离物之前已使用 BACTEC MGIT 960 系统(美国 BD 公司)进行了一线药物药敏试验,并已保存。H37Rv 标准菌株和已知对 RIF 和 INH 耐药的外部质量控制菌株(IDT3)被用作本研究的质量控制分离株。作为一种新的检测方法,检测利福平和异烟肼耐药性的毛细管法与标准的 BACTEC MGIT 960 系统进行了比较。在检测利福平和异烟肼耐药性时,毛细管法与参照法相比,灵敏度分别为 85% 和 80%;但研究发现,这两种药物的特异性值(分别为 25% 和 45.5%)较低。毛细管法检测耐药性的时间为 4-9 天不等。毛细管法是专门为快速鉴定和处理耐多药的分离株而开发的,它能在短时间内检测出耐药菌株,灵敏度相对较高,但特异性很低。我们认为,继续进行更多样本的研究,甚至通过直接从临床样本中进行研究来改进该方法将是有益的。
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引用次数: 0
[Evaluation of Viral Subgenomic RNAs and Antigen Presence in SARS-CoV-2 PCR Positive Cases]. [评估 SARS-CoV-2 PCR 阳性病例的病毒亚基因组 RNA 和抗原存在情况]。
IF 1.1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-07-01 DOI: 10.5578/mb.20240037
Kazım Batıhan Büyükzengin, Alper Akçalı, Sevil Alkan, Gökhan Akdur

Polymerase chain reaction (PCR) and antigen test (AgT) are frequently used in the diagnosis of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Routine PCR tests that detect the virus genome cannot determine whether the virus is infectious or not. However, detection of subgenomic RNA (sgRNA) produced during the replication period may indicate active viral infection. Active virus detection can offer various health and economic benefits from isolation time to treatment. Antigen tests are also considered as indicators of infectiousness since they can detect viruses above a certain load amount. The aim of this study was to use two different subgenomic RNAs and antigen test instead of genomic RNA to examine the relationship with each other and the clinic in terms of infectiousness. Evaluating the antigen test together with subgenomic RNA as an indicator of infectiousness may show the importance of this test. SARS-CoV-2 PCR positive 109 naso/oropharyngeal swab samples stored at -80 °C were included in the study. In order to confirm the PCR positivity of these samples, E gene PCR was performed and AgT, and E and N sgRNA quantitative real-time reverse transcription-PCR (RT-qPCR) detection was performed. Of the 109 SARSCoV-2 PCR positive samples, 83 (76.14%) had antigen test positivity, 88 (80.73%) had E gene sgRNA, 96 (88.07%) had N gene sgRNA and 97 (89%) had at least one sgRNA positivity.The antigen test was found positive in 77.3% of the samples in which at least one sgRNA was detected and in 66.7% of the negative samples and this difference was not statistically significant (p= 0.475). The difference between E sgRNA and AgT positivity was significant (p= 0.023). N sgRNA was positive in 98.9% of E sgRNA positive samples and 42.9% of the negative samples and this difference was statistically significant (p= 0.0001). The AgT positivity rate was found to be 98.15% (53/54) for cycle threshold (Ct) value ≤ 25, 57.14% (12/21) for Ct 25-30, and 52.94% (18/34) for Ct ≥ 30. The difference in antigen test positivity between E gRNA Ct value ≤ 25 and > 25, ≤ 29 and > 29, < 30 and ≥ 30 was statistically significant (p= 0.0001). Antigen test positivity appears to be associated with viral load and infectivity, as expected. In our study, it has been shown that sgRNAs and AgT which are indicators of infectiousness can be detected at least 10 days after the symptom period. Using these two tests together could detect infective individuals with higher accuracy and shorten the duration of hospital stay and isolation.

聚合酶链反应(PCR)和抗原检测(AgT)是诊断严重急性呼吸系统综合征冠状病毒-2(SARS-CoV-2)的常用方法。检测病毒基因组的常规 PCR 检测不能确定病毒是否具有传染性。然而,检测复制期间产生的亚基因组 RNA(sgRNA)可能表明病毒感染活跃。主动检测病毒可提供从隔离时间到治疗的各种健康和经济效益。抗原检测也被认为是传染性的指标,因为它们能检测出超过一定负荷量的病毒。本研究的目的是使用两种不同的亚基因组 RNA 和抗原检测来代替基因组 RNA,以研究它们之间的关系以及临床感染性。将抗原检测与亚基因组 RNA 一起作为传染性指标进行评估,可能会显示出该检测的重要性。本研究采集了在零下 80 摄氏度保存的 109 份 SARS-CoV-2 PCR 阳性鼻/咽拭子样本。为了确认这些样本的 PCR 阳性,进行了 E 基因 PCR 和 AgT 检测,以及 E 和 N sgRNA 实时反转录定量 PCR(RT-qPCR)检测。在 109 份 SARSCoV-2 PCR 阳性样本中,83 份(76.14%)抗原检测呈阳性,88 份(80.73%)E 基因 sgRNA 呈阳性,96 份(88.07%)N 基因 sgRNA 呈阳性,97 份(89%)至少有一个 sgRNA 呈阳性。E sgRNA 和 AgT 阳性之间的差异显著(p= 0.023)。98.9% 的 E sgRNA 阳性样本和 42.9% 的阴性样本中 N sgRNA 呈阳性,这一差异有统计学意义(p= 0.0001)。周期阈值(Ct)≤25 时,AgT 阳性率为 98.15%(53/54);Ct 25-30 时,AgT 阳性率为 57.14%(12/21);Ct ≥ 30 时,AgT 阳性率为 52.94%(18/34)。E gRNA Ct 值≤25 和>25、≤29 和>29、<30 和≥30 之间的抗原检测阳性率差异有统计学意义(p= 0.0001)。正如预期的那样,抗原检测阳性似乎与病毒载量和感染性有关。我们的研究表明,作为传染性指标的 sgRNAs 和 AgT 可在症状期后至少 10 天检测到。同时使用这两种检测方法可以更准确地检测出感染者,缩短住院时间和隔离时间。
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引用次数: 0
[Comparison of Three Different Methods in Investigating the Molecular Epidemiology of Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates]. [比较三种不同方法在调查耐碳青霉烯类鲍曼不动杆菌临床分离物分子流行病学中的应用]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-04-01 DOI: 10.5578/mb.202498131
Gülşen Uluçam Atay, Gülçin Bayramoğlu, İlknur Tosun, Ülkü Ünsal
<p><p>The aim of the study was to evaluate the relationship between carbapenem-resistant Acinetobacter baumannii isolates carrying oxacillinase-type carbapenemase genes with "international high-risk clones" (IC I, II, and III) by different molecular epidemiological methods and to statistically compare the concordance and discrimination power of the methods. Carbapenem-resistant and moderately susceptible A.baumannii isolates from non-repeating blood cultures of 72 patients were included in the study. The presence of "blaOXA-23 , blaOXA-24 , blaOXA-51 ve blaOXA-58 " genes within OXA-type carbapenemases was detected by polymerase chain reaction (PCR) method and confirmed by DNA sequence analysis. Pulsed f ield gel electrophoresis (PFGE), multilocus sequence typing (MLST) and matrix-assisted laser desorption/ ionization time- of-flight mass spectrometry (MALDI-TOF MS) analyses were performed to evaluate the clonal relations of IC I, II and III clones together with clinical isolates. In the statistical comparison of the methods, discrimination power was evaluated by Simpson index of diversity (SID) and concordance by "Wallace coefficient". All of the isolates were found to carry blaOXA-23 and blaOXA-51 genes. As a result of the bioinformatic analysis of the four isolates selected for sequence analysis; blaOXA-23 and blaOXA-51 genes were detected in the selected isolates, and the analysis of two isolates carrying blaOXA-51 gene showed 99% similarity with blaOXA-92 gene. The isolates were clustered into five pulsotypes (A, B, C, D and E) according to ≥ 85% similarity coefficient by PFGE. The isolates and RUH 875, RUH 134, LUH 5875 strains belonging to high-risk clones ICI, ICII and ICIII, respectively, were divided into five main groups [A (n= 58), B (n= 8), C (n= 4), D (n= 4) and E (n= 1)] and 10 subgroups (A1, A2, A4, A5, A6, A9, B1, B4, C3, D1) by PFGE. IC clone III (E1) and seven strains showed singleton PFGE profiles (A3, A7, A8, B2, B3, C1, C2). ICII was found in A5 subtype, ICI in C1 subtype and ICIII in E1 subtype. By PFGE subtype groups, 18 pulsotypes were determined and ST1, ST2, ST81, ST157 and ST604 sequence types were found in 20 isolates randomly selected from pulsotypes according to MLST Pasteur scheme (cpn60, fusA, gltA, pyrG, recA, rplB, rpoB). Principal component analysis (PCA) of the spectra of 72 A. baumannii isolates and ICI, ICII and ICIII clones was performed by MALDI-TOF MS. In PCA analysis, the cluster distance level was defined as 1.5 and the isolates were divided into three clusters. IC clone I, II and III together with 70 clinical isolates were grouped in one cluster, while two clinical isolates (AB083 and AB0115) formed singleton clusters. There was no significant agreement between MALDI-TOF MS; MLST and PFGE data according to Wallace coefficient. It was found that PFGE method gave significant results in terms of discrimination power with SID coefficient, MALDI-TOF MS PCA analysis had the lowest discrimination power value,
该研究旨在通过不同的分子流行病学方法评估携带氧青霉素酶型碳青霉烯酶基因的耐碳青霉烯类鲍曼不动杆菌分离株与 "国际高风险克隆"(IC I、II 和 III)之间的关系,并对这些方法的一致性和鉴别力进行统计比较。研究纳入了从 72 名患者的非重复血液培养物中分离出的耐碳青霉烯类和中度易感鲍曼不动杆菌。通过聚合酶链反应(PCR)方法检测了 OXA 型碳青霉烯酶中 "blaOXA-23、blaOXA-24、blaOXA-51 和 blaOXA-58" 基因的存在,并通过 DNA 序列分析进行了确认。通过脉冲场凝胶电泳(PFGE)、多焦点序列分型(MLST)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)分析,评估了IC I、II和III克隆与临床分离株的克隆关系。在对这些方法进行统计比较时,用辛普森多样性指数(SID)评估了区分能力,用 "华莱士系数 "评估了一致性。发现所有分离株都携带 blaOXA-23 和 blaOXA-51 基因。对所选的 4 个分离株进行生物信息学分析的结果显示,在所选分离株中检测到 blaOXA-23 和 blaOXA-51 基因,对两个携带 blaOXA-51 基因的分离株的分析表明,它们与 blaOXA-92 基因的相似度为 99%。通过 PFGE 分析,根据≥85%的相似系数,将分离物分为五个脉冲型(A、B、C、D 和 E)。通过 PFGE 将分离株和分别属于高风险克隆 ICI、ICII 和 ICIII 的 RUH 875、RUH 134 和 LUH 5875 株分为五大组[A(n= 58)、B(n= 8)、C(n= 4)、D(n= 4)和 E(n= 1)]和 10 个亚组(A1、A2、A4、A5、A6、A9、B1、B4、C3、D1)。集成电路克隆 III(E1)和 7 个菌株(A3、A7、A8、B2、B3、C1、C2)的 PFGE 图谱显示为单一。在 A5 亚型中发现了 ICII,在 C1 亚型中发现了 ICI,在 E1 亚型中发现了 ICIII。通过 PFGE 亚型分组,确定了 18 个脉冲型,并根据 MLST 巴氏方案(cpn60、fusA、gltA、PYRG、recA、rplB、rpoB)从脉冲型中随机选择了 20 个分离株,发现了 ST1、ST2、ST81、ST157 和 ST604 序列类型。利用 MALDI-TOF MS 对 72 个鲍曼尼氏菌分离物和 ICI、ICII 和 ICIII 克隆的光谱进行了主成分分析(PCA)。在 PCA 分析中,聚类距离水平被定义为 1.5,分离物被分为三个聚类。集成电路克隆 I、II 和 III 与 70 个临床分离物组成一个聚类,而两个临床分离物(AB083 和 AB0115)组成单个聚类。根据华莱士系数,MALDI-TOF MS、MLST 和 PFGE 数据之间没有明显的一致性。研究发现,PFGE 方法在 SID 系数的鉴别力方面效果显著,MALDI-TOF MS PCA 分析的鉴别力值最低,而 PFGE 和 MLST 的 Wallace 系数结果一致。总之,MALDI-TOF MS 可能无法像 PFGE 和 MLST 一样作为鲍曼不动杆菌流行病学分析的金标准方法,MALDI-TOF MS 所使用的流行病学分型方案需要改进和发展。
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