非结核分枝杆菌耐药情况分析

目的探讨非结核分枝杆菌(non-tuberculosis mycobacterium,以下简称NTM)的流行病学特征,观察和分析NTM的耐药情况,为NTM肺病的防治策略提供科学依据。方法对2006～2010年期间收集的分枝杆菌进行药物敏感性测定、菌种鉴定,对鉴定为NTM的病例进行综合分析。结果 5年间对765个菌株进行了结核分枝杆菌和非结核分枝杆菌的菌型鉴,确诊为NTM的为79株(10.32%),其中77株对异烟肼、对氨基水杨酸钠、乙胺丁醇、利福平、链霉素等常用抗结核药物存在不同程度的耐药,耐药率高达97.46%,且大多菌株同时对多种抗结核药物耐药。结论 NTM对抗结核药物具极高的耐药性。为此,对可疑肺结核患者应尽可能进行痰分离培养和多种抗结核药物敏感性测定。研究、探索抗NTM的新药及更为有效的诊断方法是当务之急。     

54株分枝杆菌国际标准株16S-23S rRNA转录间隔区序列分析

目的分析54株分枝杆菌国际标准株16S-23SrRNA转录间隔区(ITS)序列,为临床分离株的鉴定提供参考。方法用16S-23SrRNA ITS序列分析法对54株分枝杆菌国际标准株进行分析,构建系统发育树,计算相似性百分比。结果除灰尘与微黄分枝杆菌;田野与千田分枝杆菌;抗热与副偶然分枝杆菌,奥布与母牛结核分枝杆菌;杜氏与猪分枝杆菌;金色与东海分枝杆菌,海与溃疡分枝杆菌、科莫斯分枝杆菌;2株结核分枝杆菌与田鼠分枝杆菌、非洲分枝杆菌的16S-23SrRNA ITS序列完全相同无法鉴别外,其他各分枝杆菌菌种间16S-23SrRNAITS序列均不相同,可以得到很好的鉴别。结论 16S-23SrRNA ITS序列分析是一种很好的鉴定分枝杆菌的方法,国际标准株16S-23SrRNA ITS序列的研究弥补了基因数据库的不足。     

Radical surgical debridement alone for treatment of carpal tunnel syndrome caused by mycobacterium avium complex flexor tenosynovitis: case report

We present a case of proliferative flexor tenosynovitis caused by Mycobacterium avium complex resulting in recurrent symptoms of median nerve compression. The patient was treated with radical tenosynovectomy and release of the carpal tunnel without antimicrobial therapy. She remains symptom-free and is apparently disease-free more than 2 years after surgery.     

Mycobacterium fortuitum-induced persistent parotitis: successful therapy with clarithromycin and ciprofloxacin

BACKGROUND: Parotitis caused by nontuberculous mycobacteria, a very rare disease entity, has never been reported to be caused by Mycobacterium fortuitum (M. fortuitum) in the literature. METHODS AND RESULTS: An 8-year-old girl was seen with painful swelling of the right parotid gland despite antibiotic treatment of more than 1 month. Elevated serum amylase activity and diffuse contrast-enhanced CT of the parotid gland confirmed the diagnosis of parotitis. Histopathological study of specimens taken from the right parotid tail mass showed granulomatous inflammation with acid-fast positive bacilli; culture later confirmed M. fortuitum. After administration of clarithromycin and ciprofloxacin for 9 consecutive months, the parotitis and parotid tail mass were completely resolved at follow-up examination. CONCLUSION: To our knowledge, this is the first case report of parotitis caused by M. fortuitum and its successful medical treatment.     

74例结核性淋巴结炎针吸活检标本结核菌培养临床分析

目的本研究对临床怀疑为结核性淋巴结炎的病人采用细针活检标本做细菌学培养,评价其诊断价值及耐药发生率。方法对110例临床怀疑为结核性淋巴结炎病人采用细针针吸活检术,对所获病理组织进行细胞组织学和细菌培养检查。结果74例患者经组织细胞学及细菌学被诊断为结核性淋巴结炎,其中14例（18．9％）吸引标本分离培养到结核分枝杆菌。14例病人中12例为人型结核分枝杆菌,2例牛型结核分枝杆菌。大多对四种常规抗结核药物敏感,4例（28．5％）一种或多种药物耐药。结论对临床怀疑为结核性淋巴结炎者,凡窦道、脓肿型或见有脓性吸引标本都应作结核菌培养,而结节型培养结果均为阴性,临床意义不大。     

HLA Antigens and Nontuberculous Mycobacterial Lung Disease in Korean Patients

Background  Human leukocyte antigen (HLA) molecules are known to play an important role in host-defense mechanisms. The aim of this study was to evaluate the association between HLA alleles and lung disease caused by nontuberculous mycobacteria (NTM) in Korean patients. Methods  Seventy-eight patients with NTM lung disease (48 patients with Mycobacterium avium-intracellulare complex [MAC] infection and 30 patients with Mycobacterium abscessus infection) were included in the study. HLA-A, -B, and -DRB1 genotyping was performed by polymerase chain reaction using sequence-specific primers. Data from 485 healthy Korean individuals were used as a control. Results  When compared to controls, patients with NTM lung disease showed an increased frequency of DRB1*11 (OR = 1.91, 95% confidence interval [CI] = 1.01–3.64, p = 0.045, corrected p [pC] > 0.05). In the subgroup analysis, patients with MAC lung disease had an increased frequency of B*46 (OR = 2.23, 95% CI = 1.05–4.73, p = 0.044, pC > 0.05). Conclusions  Our data suggest that in a Korean population, patients with NTM lung disease and healthy subjects differ in the frequencies of some HLA alleles. However, when considering corrected p values, our findings are inconclusive. S.-W. Um and C.-S. Ki contributed equally to this work.     

新型大肠杆菌-分枝杆菌穿梭载体的构建及卡那霉素抗性基因表达的研究

用 PCR（polymerase chain reaction）技术克隆了卡介苗（Bacille Calmette-Guerin,BCG）热休克蛋白 65（heat shock Protein 65, hsp65）启动子序列,构建成穿梭载体ME-65。该载体含有分枝杆菌和大肠杆菌的质粒复制基因、BCG的hsp65高效启动子序列以及在分枝杆菌和大肠杆菌中都能表达的卡那霉素抗性基因。将其转化到耻垢分枝杆菌和BCG中均可稳定复制并表达卡那霉素抗性。质粒对宿主菌的生长无明显的影响,未观察到质粒的丢失和突变。该载体具有分子量小、容量大、转化效率高、复制稳定、含有多克隆位点等优点,将外源基因插入到hsp65启动子的下游,可望在BCG中获得高效表达。为BCG和其它分枝杆菌发展成多价重组疫苗开辟了新途径。     

The Mycobacteriophage Ms6 LysB N-Terminus Displays Peptidoglycan Binding Affinity

Double-stranded DNA bacteriophages end their lytic cycle by disrupting the host cell envelope, which allows the release of the virion progeny. Each phage must synthesize lysis proteins that target each cell barrier to phage release. In addition to holins, which permeabilize the cytoplasmic membrane, and endolysins, which disrupt the peptidoglycan (PG), mycobacteriophages synthesize a specific lysis protein, LysB, capable of detaching the outer membrane from the complex cell wall of mycobacteria. The family of LysB proteins is highly diverse, with many members presenting an extended N-terminus. The N-terminal region of mycobacteriophage Ms6 LysB shows structural similarity to the PG-binding domain (PGBD) of the φKZ endolysin. A fusion of this region with enhanced green fluorescent protein (Ms6LysBPGBD-EGFP) was shown to bind to Mycobacterium smegmatis, Mycobacterium vaccae, Mycobacterium bovis BGC and Mycobacterium tuberculosis H37Ra cells pretreated with SDS or Ms6 LysB. In pulldown assays, we demonstrate that Ms6 LysB and Ms6LysBPGBD-EGFP bind to purified peptidoglycan of M. smegmatis, Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis, demonstrating affinity to PG of the A1γ chemotype. An infection assay with an Ms6 mutant producing a truncated version of LysB lacking the first 90 amino acids resulted in an abrupt lysis. These results clearly demonstrate that the N-terminus of Ms6 LysB binds to the PG.     

河南省尉氏县结核病的病原学调查及结核分枝杆菌耐药性分析

目的 调查河南省结核病的分枝杆菌菌种类型、分布特征及耐药状况。 方法 对研究现场所有疑似结核病患者进行痰涂片和痰菌培养;用鉴别培养基和双重聚合酶链反应(PCR)方法对分离的菌株进行菌种鉴定,用比例法进行药物敏感性检测。 结果 共收集患者2661份痰标本,应用痰涂片、痰培养和双重PCR三种方法联合应用进行病原学分析,发现涂片阳性者179份(占6.5%);培养阳性菌株245份,阳性率为9.2%,其中225株为结核分枝杆菌,占91.8%,结核分枝杆菌复合群6株,占2.4%,13株为非结核分枝杆菌(NTM),占5.3%,1株为结核分枝杆菌与非结核分枝杆菌混合感染,占0.4%。药敏检测显示结核分枝杆菌总体耐药率为24.0%(54/225),4种药物的耐药率链霉素16.9%(38/225),异烟肼 16.0%(36/225),利福平 9.8%(22/225),乙胺丁醇8.4%(19/225)。耐单药菌株20株,占8.9% ,耐2种药以上的多药耐药为15.1%(34/225),其中,同时耐异烟肼和利福平及2种以上抗生素菌株即耐多药株(MDR-TB)菌株19株,占8.4%(19/225)%。男、女性结核病患者耐药率差异无统计学意义(2=2.8214,P0.05),耐药与年龄的相关性差异有统计学意义(2=8.3868,P0.05),复治病例耐药率与初治病例耐药率差异有统计学意义(2=13.7193,P0.05);复治病例MDR-TB耐药率与初治病例MDR-TB耐药率差异有统计学意义(211.1361,P0.05)。 结论 河南省尉氏县的结核病病原菌包括结核分枝杆菌、结核分枝杆菌复合群及非结核分枝杆菌,耐药情况较为严重,尤其对复治病例、较年轻病例应引起重视。