BioVector® ATCC 33559 结肠弯曲杆菌标准质控株

BioVector® ATCC 33559 Campylobacter coli Quality Control Strain

第一部分 中文说明

一 产品基本信息与遗传学背景

• 菌种名称 BioVector® ATCC 33559 结肠弯曲杆菌（大肠弯曲杆菌）

• 别名与同义词 Campylobacter coli (Doyle) Veron and Chatelain；同型异名 Vibrio coli。

• 分类学地位 细菌域（Bacteria），变形菌门（Pseudomonadota），弯曲菌属（Campylobacter）。

• 保藏机构菌株号 ATCC 33559 = CIP 70.80 = DSM 4689 = NCTC 11366。

• 分离来源与菌株性质

  • 原始分离自猪粪便（Pig feces）。

  • 该菌株是经国际公认的全基因组测序标准模式物种菌株（Type strain）。

• 生物安全级别 2级（Risk Group 2，主要引起食源性胃肠炎）。

二 生理生化特征与特殊培养环境

• 形态学特征 显微镜下表现为革兰氏阴性（Gram-negative）、能运动、呈细长弯曲或微螺旋状、逗点状的杆菌。

• 气体生长环境 属于严格的微需氧菌（Microaerophilic）。在进行体外人工培养时，必须在微需氧（通常为 5% 氧气、10% 二氧化碳、85% 氮气）的气体生成包或厌氧罐中孵育。

• 推荐标准培养基

  • 液体基础 ATCC 富集肉汤培养基（如 #1115 培养基）。

  • 固体平板 弯曲菌选择性血琼脂平板（如 #260 琼脂平板或巧克力琼脂平板）。

• 物理培养参数 37摄氏度恒温培养 48到72小时（所有试管或斜面在微需氧罐中应保持瓶盖松开状态以利于气体交换）。

三 冻干菌种复苏与传代标准步骤

1. 菌株复苏操作

   • 无菌条件下打开包含冻干菌种的西林瓶。

   • 用无菌吸管吸取约 0.5 到 1.0 mL 预热的完全肉汤培养基，注入冻干管中，轻轻吹吸使菌块完全溶解重悬。

   • 将溶解后的菌悬液全部接种到 5 到 6 mL 的液体富集肉汤试管中，同时吸取数滴菌液涂布接种到血琼脂平板或琼脂斜面上。

2. 培养与保藏

   • 迅速将接种完成的试管和平板放进微需氧装置中，37摄氏度孵育 48小时。

   • 注：复苏后的前48小时内，切勿频繁打开密封培养皿或厌氧罐，以避免因氧气浓度波动导致脆弱的弯曲菌死亡。

   • 长期保藏 推荐使用含有 10% 灭菌甘油的保护剂，置于液氮或 -80摄氏度超低温冰箱中冷冻保存。

四 核心科研与工业应用方向

1. 微生物实验室质量控制（QC Organism） BioVector® ATCC 33559 作为标准的弯曲菌模式菌株，被广泛用作商业化培养基性能测试、革兰氏染色试剂质控、弯曲菌微量生化鉴定试剂盒（如 API 系统）的阳性对照标准株。

2. 食品安全与肠道传染病（食源性致病菌）研究 用于建立针对食品（如家禽肉类、畜产品）中结肠弯曲杆菌残留的标准前增殖、富集检测方法（依据 FDA BAM 或 ISO 11133 标准规范）。

3. 分子诊断与基因扩增检测标准品 该菌株提取的定量基因组 DNA（如 33559DQ 分子标准品）作为金标准线，广泛用于多重实时荧光定量 PCR（qPCR）、下一代测序（NGS）等临床分子诊断测定法的研发、验证与 Lot-to-Lot 稳定性监控。

PART 2 ENGLISH SECTION

I General Information and Genetic Background

• Culture Name BioVector® ATCC 33559 Campylobacter coli (Doyle) Veron and Chatelain

• Synonyms Homotypic synonym Vibrio coli Doyle 1948.

• Taxonomic Classification Domain Bacteria, Phylum Pseudomonadota, Genus Campylobacter, Species Campylobacter coli.

• Repositories Identifiers ATCC 33559 = CIP 70.80 = DSM 4689 = NCTC 11366.

• Isolate History and Strain Nature

  • Originally isolated from pig feces.

  • Serves as a whole-genome sequenced bacterial type strain recognized globally.

• Biosafety Level BSL-2 (Risk Group 2 due to its enteric pathogenicity).

II Morphological Attributes and Cultivation Media

• Morphology Reveals classic Gram-negative, motile, curved, comma-shaped, or slender helical rod profiles under oil-immersion microscopy.

• Growth Atmosphere Requires strict microaerophilic environments to proliferate. Standard culture should occur under controlled gas layers (ideally 5% O2, 10% CO2, and 85% N2).

• Standard Growth Media Recipes

  • Basal Liquid Broth ATCC Enrichment Medium (e.g., #1115 Formulation).

  • Solid Agar Campylobacter selective Blood Agar plates (or Chocolate Agar formulations).

• Physical Incubation Thresholds Incubated closely at 37 degrees Celsius for 48 to 72 hours, maintaining loose caps inside microaerophilic jars to ensure seamless gas exchange tracks.

III Rehydration and Maintenance Protocols

1. Standard Pelleted Culture Recovery

   • Aseptically unpack the lyophilized vial inside a certified bio-safety hood.

   • Rehydrate the dried disc pellet by injecting 0.5 to 1.0 mL of pre-warmed broth base directly into the ampoule, mixing gently until thoroughly dissolved.

   • Inoculate the mixture into a secondary test tube carrying 5 to 6 mL of fresh liquid medium and streak several drops across blood agar plates or slants.

2. Continuous Preservation Parameters

   • Promptly deposit the fresh culture matrix inside a microaerophilic container and run incubation at 37 degrees Celsius for 48 hours.

   • Tip: Avoid opening the closed agar plates or venting the anaerobic jar during the preliminary 48-hour cycle to shield sensitive cells from oxygen shock.

   • Long-term Storage Cryopreservation is achieved at liquid nitrogen thresholds or in a -80 degrees Celsius freezer using 10% sterile glycerol cryoprotectant arrays.

IV Strategic Research and Quality Control Applications

1. Microbiology Laboratory Quality Control (QC Organism Reference) BioVector® ATCC 33559 is widely used for media performance evaluations, validating Gram stain accuracy, confirming identification kits, and auditing standard clinical bacteriological procedures.

2. Food Safety and Enteric Pathogen Screening Deployed to standardise enrichment metrics and validate pathogen screening methods from meat matrices following official FDA BAM or ISO 11133 compliance rules.

3. Molecular Standards Development and Diagnostic Assays Purified quantitative genomic DNA derived from this specific clone (e.g., ATCC 33559DQ standards) is utilized to generate exact standard curves for qPCR or validate NGS pipelines.

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