研究与开发杂志
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国际标准期刊号: 2311-3278
国际标准期刊号: 2311-3278
费哈特·朱迪
结核病是一种非常古老的传染病。这种病理学的病原体是结核分枝杆菌，由罗伯特·科赫 (Robert Koch) 在 19 世纪发现。结核杆菌感染了第一批原始人类，并与主题共同进化。这种感染通常始于吸入病人排出的受污染的飞沫，最低感染剂量非常低，从 1 到 10 个杆菌不等。据世界卫生组织（WHO）称，世界上近三分之一的人口感染了结核杆菌。2015年，87%的新病例发生在30个结核病高负担国家。六个国家占新增病例的 60%：印度、印度尼西亚、中国、尼日利亚、巴基斯坦和南非。这种疾病的发病率平均下降了1。 研究目的 本研究的目的是验证 BACTEC MGIT 960 在阿尔及利亚巴斯德研究所结核病和分枝杆菌科与 LJ 培养基上的经典培养相比在肺结核诊断中的贡献。 材料与方法： 设置和道德考虑： 阿尔及尔巴斯德研究所（IPA）结核病和分枝杆菌实验室在阿尔及利亚的结核病防治工作中占有重要地位，除了是诊断和抗结核药物检测的国家参考实验室外，还参与监督、监测和报告参与结核病诊断的整个国家实验室网络的结果。它也是与世界卫生组织在非洲地区合作的超国家实验室。 Oral consents were obtained from all patients prior to specimen’s collection, and ethical considerations were taken into account during all steps of the study. The patient’s data and results were maintained in secure database. Materials and procedures Three types of pulmonary samples were included in the study: expectoration, gastric tubing and bronchial aspiration. The collection of these samples was done in clean spittoons. Each sample sent to the laboratory was accompanied with information sheet of the patient. The samples were stored at + 4°C and Z-N staining were directly performed. Poly-microbial samples subjected to prior decontamination before they were cultured, and Petroff’s decontamination technique was used, without neutralization. For each sample, 2 tubes of Lowenstein-Jensen were inoculated. Before inoculating the MGIT tubes, samples were processed using the BBLMycoPrep kit containing a mixture of N-acetyl-L-cysteine and 2% sodium hydroxide (NALC-NaOH), following the Kubica’s Protocol. Positive tubes in BACTEC MGIT 960 were systematically inoculated on Lowenstein-Jensen media and Ziehl-Neelsen staining was performed for each tube. Finally, a rapid identification with immuno-chromatoghaphic TBC ID was applied. Antibiogram on solid medium, proportions method: The colonies (about 1 mg) are placed in a 50 ml flask containing glass beads, stirred for 10 min, in order to dissociate the colonies. From the initial suspension and dilution 10-2, two L-J tubes containing, each, one of the first line anti-tuberculosis drugs: Isoniazid (0.2 mg/ ml), streptomycin (4 mg/ml), rifampicin (40 mg/ml) and ethambutol (2 mg/ml), were inoculated. For each sample, two single L-J tubes (without antibiotics) were inoculated as controls. L-J containing specific media tubes (TCH, PNB and PAS), for confirmation of identification, were inoculated with the initial suspension. First verification of tubes, after incubation at 37°C, was done the 28th day, a second and final lecture at the 42nd day. After counting of colonies number on both types of tubes, a proportion ratio between the number of colonies with antibiotics and the number of colonies on the control tubes is expressed as a percentage. Below 1% “critical proportion”, the strain is sensitive, above or equal to 1%, it was considered as resistant. MDRs isolates were submitted to a second antibiogram, against Ofloxacin and Kanamycin, to verify the existence of XDR isolates. Antibiogram on liquid medium, BACTEC MGIT 960: The first day of the positivity of a MGIT culture tube is considered as day “D-0”. Inocula for this antibiogram should be prepared between “D-1” and “D-5”. Five tubes were labeled for each isolate to be tested, a growth control tube and 4 tubes containing the antibiotics; streptomycin (1 μg/ml), Isoniazid (0.1 μg/ml), rifampicin (1 μg/ml), ethambutol (5 μg/ml). Reading is done automatically every 60 minutes, the sensitivity test is recorded between 5 to 12 days, and the result is obtained in the form of a printed report. Statistical Analysis: 进行统计分析以计算频率和显着性差异，在适当时使用单向方差分析检验或Kruskall-Wallis，或分别通过卡方检验和费舍尔精确检验。使用列联表评估所考虑的变量之间的关联，并且所有报告的 p 值都是双边的。p < 0.05 被认为是显着的。 结果与讨论： 在过去的几十年里，结核病感染的发病率显着上升。显然，该病有利因素的增多以及多重耐药菌株的出现，无论是发达国家还是发展中国家，都是国际社会面临的重大健康问题。这种令人震惊的情况促使人们研究和开发更快速、更有效的检测和治疗手段，以降低死亡率。除其他外，与这种疾病的竞赛是合理的，因为它是 2014 年艾滋病之前最致命的疾病。一些研究表明，液体和固体介质的结合是诊断艾滋病毒的基本原则。结核。然而，在大多数非洲实验室，由于液体介质成本高昂，这种组合很少可用。因此，仅使用固体培养基（LJ）进行培养和药敏试验。 注：这项工作部分在2018年10月11-12日在俄罗斯莫斯科举行的第21届欧洲生物技术大会上展示