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首页> 外文OA文献 >Application of the C18-Carboxypropylbetaine Specimen Processing Method to Recovery of Mycobacterium avium subsp. paratuberculosis from Ruminant Tissue Specimens
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Application of the C18-Carboxypropylbetaine Specimen Processing Method to Recovery of Mycobacterium avium subsp. paratuberculosis from Ruminant Tissue Specimens

机译：C18-羧丙基甜菜碱样品处理方法在鸟分枝杆菌亚种回收中的应用。反刍动物组织标本中的副结核病

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摘要

The causative agent of Johne's disease is Mycobacterium avium subsp. paratuberculosis. This is a chronic, debilitating gastrointestinal disorder that affects ruminants and is responsible for significant economic loss. The specimen processing method that combines C18-carboxypropylbetaine (CB-18) treatment and lytic enzyme decontamination has been shown to improve the diagnosis of mycobacterioses. This processing method was applied to the isolation of M. avium subsp. paratuberculosis from ruminant tissue samples. The BACTEC 12B liquid culture system was used but was supplemented with 1% egg yolk emulsion, 4 μg of mycobactin J, and 0.5% pyruvate (12B/EMP) for use in conjunction with this method. The final concentration of antibiotics used was 10 μg of vancomycin, 30 μg of amphotericin B, and 20 μg of nalidixic acid (VAN) per ml. A 7H10-based solid medium was also used that included mycobactin J, pyruvate, and VAN but excluded the egg yolk emulsion (7H10/MPV). Several M. avium subsp. paratuberculosis isolates were examined during the evaluation of this processing method. It was observed that treatment with lytic enzymes stimulated the growth of M. avium subsp. paratuberculosis; however, the growth of one isolate was markedly inhibited due to the presence of vancomycin. Subsequently, the vancomycin concentration in the VAN formulation was reduced to 2 μg/ml. A blinded panel of 60 previously characterized tissue samples from bovine and bison were then processed and analyzed by smear and culture. Historically, 31 and 37 specimens were classified as positive by histology and culture, respectively. The overall sensitivity and specificity of smear relative to culture following CB-18 processing were 97.6 and 89.5%, respectively. The 12B/EMP/VAN liquid culture system recovered M. avium subsp. paratuberculosis from 39 specimens, whereas 7H10/MPV and Herrold's egg yolk media recovered M. avium subsp. paratuberculosis from 26 and 16 specimens, respectively. The average times to positive were 7.4 ± 8.3, 29.9 ± 2.6, and 24 ± 0 days, respectively. The contamination rates were 4.8, 22.6, and 20.0%, respectively.
机译：约翰氏病的病原体是鸟分枝杆菌亚种。副结核病。这是一种慢性的，使人衰弱的胃肠道疾病，会影响反刍动物并造成重大的经济损失。结合C18-羧丙基甜菜碱（CB-18）处理和裂解酶去污的标本处理方法已被证明可以改善分枝杆菌的诊断。该处理方法被应用于鸟分枝杆菌亚种的分离。反刍动物组织样本中的副结核病。使用了BACTEC 12B液体培养系统，但补充了1％的蛋黄乳剂，4μg的霉菌素J和0.5％的丙酮酸（12B / EMP），可与该方法结合使用。最终使用的抗生素浓度为每毫升10μg万古霉素，30μg两性霉素B和20μg萘啶酸（VAN）。还使用了基于7H10的固体培养基，其中包括霉菌素J，丙酮酸和VAN，但不包括蛋黄乳剂（7H10 / MPV）。几个鸟分枝杆菌亚种。在评估该加工方法的过程中检查了副结核菌分离株。观察到用裂解酶处理刺激了鸟分枝杆菌亚种的生长。肺结核然而，由于存在万古霉素，一种分离株的生长受到明显抑制。随后，将VAN制剂中的万古霉素浓度降低至2μg/ ml。然后对来自牛和野牛的60个先前表征的组织样品进行盲法处理，并通过涂片和培养进行分析。从历史上看，分别通过组织学和文化将31个和37个样本分类为阳性。相对于CB-18处理后的培养，涂片的总体敏感性和特异性分别为97.6和89.5％。 12B / EMP / VAN液体培养系统回收了鸟分枝杆菌亚种。从39个标本中检出副结核病，而7H10 / MPV和Herrold的蛋黄培养基恢复了鸟分枝杆菌亚种。分别来自26和16个标本的肺结核。出现阳性的平均时间分别为7.4±8.3、29.9±2.6和24±0天。污染率分别为4.8％，22.6％和20.0％。

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