| 曹蕊,余萌,马仕洪.《中国药典》拟收载洋葱伯克霍尔德菌群(Bcc)检查法中标准菌株的稳定性研究[J].中国药事,2022,36(7):724-735 |
| 《中国药典》拟收载洋葱伯克霍尔德菌群(Bcc)检查法中标准菌株的稳定性研究 |
| Stability Study on Standard Strains in the Burkholderia cepacia Complex (Bcc) Inspection Method to Be Included in the Chinese Pharmacopoeia |
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| DOI：10.16153/j.1002-7777.2022.07.001 |
| 中文关键词: 洋葱伯克霍尔德菌群 16S rRNA RiboPrinter全自动基因指纹图谱鉴定系统 Biolog全自动生化鉴定系统 菌株稳定性 |
| 英文关键词: Burkholderia cepacia Complex 16S rRNA RiboPrinter automatic genetic fi ngerprint identifi cation system biolog automatic biochemical identifi cation system strain stability |
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| 中文摘要: |
| 目的：应用现有微生物分类鉴定技术手段，对《中国药典》拟收载洋葱伯克霍尔德菌群(Bcc) 检查法中所选用的标准菌株进行冻存及传代稳定性考察。方法：选取美国药典USP43-NF38 Bcc检查法中的一株Bcc标准菌株ATCC25416为对照菌株。以3株拟收载标准菌株和1株对照标准菌株为试验菌株，分别对这些试验菌株冻存0.5年、1年、2年的样本及连续6代的传代样本进行菌落表型观察、Biolog生化鉴定分析、16S rRNA基因测序及RiboPrinter全自动基因指纹图谱分析，并使用BioNumerics 7.6对核糖体图谱数据进行聚类分析。结果：试验菌株在经冻存及传代后，各菌株表型无明显差异；菌株的Biolog生化鉴定结果虽略有差异，但菌株关键生化代谢属性稳定，其鉴定结果均属Bcc；同时，各菌株的16S rRNA测序结果一致；核糖体带型相似值均大于0.95。结论：在本研究考察的冻存时间及传代次数内，3株《中国药典》洋葱伯克霍尔德菌群检查法拟收载的标准菌株的表型、基因型相对稳定。 |
| 英文摘要: |
| Objective: Using the existing methods of microbial classifi cation and identifi cation techniques to investigate the cryopreservation and passage stability of the standard strains selected in the test of Burkholderia cepacia Complex in the Chinese Pharmacopoeia. Methods: A Bcc standard strain ATCC25416 in the USP43- NF38 Bcc inspection method was selected as the control strain. Three candidate standard strains and one control strain were selected as the test strains, then the colony phenotype, Biolog biochemical identifi cation system, 16S rRNA gene sequencing and RiboPrinter automatic genetic fi ngerprinting analysis were performed on samples of these experimental strains cryopreserved for 0.5, 1 and 2 years and passage samples for 6 consecutive generations. The BioNumerics 7.6 software was used to perform the cluster analysis of the fingerprint data. Results: Subsequent to the cryopreservation or subculture of test strains of Burkholderia cepacia Complex, the phenotypes of the strains had no signifi cant diff erence. The Biolog biochemical identifi cation results of the strains were all Bcc. Although there were slight diff erences, the key biochemical metabolic properties of the strains were stable; at the same time, the 16S rRNA sequencing results were consistent and the ribotyping bands similarity values of each strain were greater than 0.95. Conclusion: Within the freezing storage time and passage times investigated in this study, the phenotypes and genotypes of the these three standard strains included in the test of Burkholderia cepacia Complex in Chinese Pharmacopoeia are relatively stable. |
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