抗结核新药普托马尼研究进展

由于对一线药物存在很大的耐药性,广泛耐药结核病和多药耐药结核病往往很难完全治愈。普托马尼（Pretomanid,PA-824）是目前临床试验评估中的治疗耐药结核分枝杆菌的优选药物之一,该药通过抑制细胞壁的形成和呼吸中毒两种机制,对复制型和静止型结核分枝杆菌均表现出显著的杀菌活性。本文通过探讨PA-824释放活性氮物质对结核分枝杆菌产生强氧化作用和细胞毒作用、延长早期杀菌活性、降低患者痰中结核分枝杆菌载量的速度、引起可控性不良反应等作用,全面了解该药物的抗结核作用机理,包括抑制结核分枝杆菌菌株耐药性的发展、缩短结核病治疗的持续时间等。本文还阐述了PA-824、贝达喹啉和利奈唑胺联合治疗,该联合方案具有相当的杀菌活性,在杀灭细菌和抑制耐药性方面具有协同或相加的作用潜力,对XDR-TB或者MDR-TB具有更好的治疗效果。有望进一步改善目前结核病的治疗现状     

荧光定量 PCR 检测结核杆菌 DNA 与涂片 抗酸染色诊断肺结核的效果比较

〔摘 要〕 目的：比较荧光定量聚合酶链式反应（PCR）检测结核杆菌脱氧核糖核酸（DNA）与涂片抗酸染色诊断肺结 核的效果。 方法：收集 2017 年 1 月至 2020 年 12 月在新乡市第一人民医院就诊的疑似肺结核患者 95 例的痰液标本 127 份, 对所有痰液标本均实施荧光定量 PCR 检测结核杆菌 DNA 及涂片抗酸染色检测,以痰培养结果为金标准,比较两种检测方 法的诊断效能。 结果：痰培养结果显示,95 例疑似肺结核患者中,61 例患者确诊,确诊率为 64.21 %。荧光定量 PCR 检测 的准确度、特异度、灵敏度及阳性预测值、阴性预测值均明显高于涂片抗酸染色检测,差异均具有统计学意义（P ＜ 0.05）。 结论：荧光定量 PCR 检测结核杆菌 DNA 对肺结核的诊断效能明显高于涂片抗酸染色检测。     

显微镜观察药物敏感性技术快速检测痰标本中结核分枝杆菌的研究

目的探讨显微镜观察药物敏感性技术(MODS)在痰液标本中结核分枝杆菌(MTB)的检测效果,以评价其临床应用价值。方法收集150份患者临床痰标本,应用MODS技术、罗氏培养法进行MTB的检测,将两法的结果进行对比分析。结果 MODS技术与罗氏培养法结果符合率为77.8%,MODS技术检测痰标本中MTB的敏感性为93.3%,特异性为70.0%,阳性预测值(PPV)为60.8%,阴性预测值(NPV)为95.4%,正确性为77.8%。结论 MODS技术检测痰标本中MTB具有快速、灵敏、简便等优点,可作为结核病细菌学快速检测的新方法之一。     

上海市普陀区1996～2003年间原发结核分枝杆菌感染的耐药性分析

目的 了解我区1996～2003年间结核分枝杆菌原发耐药性.方法 采用绝对浓度法进行异烟肼（INH）、利福平（RFP）、链霉素（SM）、乙胺丁醇（EMB）4种抗结核药物的耐受性测定.结果 总原发耐药率由1996～1997年的14.6%上升至2002～2003年的19.7%（上升了34.9%）.耐INH、SM、RFP率由1996～1997年的8.9%、9.8%、5.7%上升至2002～2003年的15.3%、10.9%、8.0%,（分别上升了71.9%、11.2%和40.4%）,耐EMB率由2.4%下降至0.7%,原发耐多药率2002～2003年为5.8%,与1996年相比,无明显上升.结论 本区结核分枝杆菌原发耐药性呈上升趋势,耐多药菌株仍占一定比例.     

Clinical analysis of pulmonary <Emphasis Type="Italic">Mycobacterium avium</Emphasis> complex disease in association with corticosteroid treatment

We studied the clinical characteristics of nine patients with pulmonary Mycobacterium avium complex disease occurring in association with corticosteroid drugs collected from our associated hospitals during the past 6 years. The average age of the nine patients was 62.2 years and the male/female ratio was 3 : 6. Regarding underlying disease, respiratory diseases existed in four of the patients and nonrespiratory diseases in the other five patients. The duration of corticosteroid treatment ranged from 5 months to 5 years, and the total dose of corticosteroid drugs ranged from 1.78 to 43.20 g. Pulmonary Mycobacterium avium complex disease was detected by clinical symptoms during corticosteroid treatment in six patients, and purified protein derivative was positive in three of eight patients tested. Radiological findings showed an infiltration shadow without cavity and bronchiectasis in the lower lung field. Microbiological examination was smear-positive in three patients, and the isolated mycobacterium was Mycobacterium intracellulare in five patients and Mycobacterium avium in four. Tolerance was shown to all antituberculous drugs, except for clarithromycin, in all patients. Although treatment including clarithromycin was performed for seven patients, the sputum conversion rate was 33% and an improved clinical effect was noted in only one patient. No change occurred in four and worsening occurred in four. Attention should be paid to the clinical symptoms and radiological findings of patients who have received corticosteroid drugs over a long period of time, because pulmonary Mycobacterium avium complex is characterized by atypical radiographic findings with no relationship to the total dose or duration of the administered corticosteroid drugs. Received: August 22, 2002 / Accepted: October 21, 2002     

Disseminated tuberculosis with paradoxical reactions caused by a Mycobacterium tuberculosis strain belonging to the Indo-Oceanic lineage: An imported case in Japan

Tuberculosis remains a major public health concern. Millions of tuberculosis cases and associated deaths have been reported worldwide. The Indo-Oceanic lineage Mycobacterium tuberculosis is common in Southeast Asia and causes extrapulmonary lesions. Only a few case studies on this lineage with genetic analysis using whole-genome sequencing have been reported in the literature. We present a case of disseminated tuberculosis, characterized by a variety of extrapulmonary lesions and paradoxical reactions, caused by the Indo-Oceanic lineage M. tuberculosis in a woman in Myanmar. A 22-year-old Burmese woman had arthritis in the right knee, with unknown aetiology, and was referred to our hospital. Computed tomography of the trunk revealed multiple nodular shadows in both lungs; swollen mediastinal lymph nodes; and small, low-density areas in the spleen. M. tuberculosis was detected in the sputum sample, joint aspirate, subcutaneous tumor, and exudate. She experienced a variety of paradoxical reactions together with aggressive tuberculosis dissemination in all areas of the body. Whole-genome sequencing of the DNA of MTB obtained from sputum and the right cervical subcutaneous abscess confirmed the Indo-Oceanic lineage of M. tuberculosis, the predominant strain in Myanmar. The Indo-Oceanic lineage M. tuberculosis causes disseminated tuberculosis all over the body including the periungual region. When patients show unusual symptoms, physicians should consider the introduction of new strains from foreign countries. Genetic analyses of the strains are recommended to define and confirm the lineages.     

北京市复治结核病患者非北京基因型菌株谱系及耐药分析

目的 了解北京市复治结核病患者非北京基因型菌株谱系及耐药情况,为复治非北京基因型结核病患者有效防治提供科学依据。方法 采用RD105基因缺失法对复治患者所分离的结核分枝杆菌（Mycobacterium tuberculosis, MTB）进行检测,筛查出非北京基因型菌株,使用可变数目串联重复序列基因分型实验分析非北京基因型菌株的遗传特征,再使用比例法药敏实验及全基因组测序分析其耐药情况。结果 134例复治结核病患者所分离的MTB中,北京基因型为 125例（93.3%）,非北京基因型为9例（6.7%）。9例非北京基因型MTB呈现9种不同的基因型,均不成簇,为独特型。9例非北京基因型MTB均为谱系4,其中,亚谱系L4.4有4株,亚谱系L4.5有5株。耐药预测结果显示,9例非北京基因型中,6例为全敏感菌株（66.7%）,1例单耐异烟肼,耐药突变位点为ahpC_c.-48G>A,1例同时耐异烟肼与乙硫异烟胺, 耐药突变位点为fabG1_c.-15C>T,1例单耐氟喹诺酮,耐药突变位点为gyrA_p.Asp94Asn与gyrA_p.Ala90Val。全基因组测序预测耐药结果与比例法药敏结果完全一致。结论 复治结核病患者非北京基因型菌株主要为谱系4,呈明显的多态性。耐药突变位点均为常见突变位点。     

结核分枝杆菌Rv2994基因重组穿梭质粒的构建与鉴定

目的构建结核分枝杆菌假想药物外排泵蛋白编码基因Rv2994的重组穿梭质粒pMV-2994,并对其进行鉴定。方法以结核分枝杆菌H37Rv基因组为模板,应用PCR技术扩增Rv2994基因编码序列,定向克隆入融合蛋白原核表达载体pGEX-1λT,获得重组表达质粒pGEX-2994,测序鉴定。酶切该基因片段,定向重组入大肠杆菌-分枝杆菌穿梭载体pMV261,构建重组质粒pMV-2994,通过限制性内切酶酶切及PCR鉴定。结果从结核分枝杆菌H37Rv株基因组DNA中扩增出的Rv2994基因与GenBank公布的序列一致,经过酶切及PCR鉴定,表明Rv2994基因成功插入了pMV261穿梭载体。结论成功构建了重组穿梭质粒pMV-2994,为进一步研究Rv2994基因功能奠定了基础。