Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH - National Research Institute, Warsaw, Poland.
Biobank Lab, Department of Oncobiology and Epigenetics, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland.
Microbiol Spectr. 2024 Jul 2;12(7):e0425923. doi: 10.1128/spectrum.04259-23. Epub 2024 May 17.
Currently, tuberculosis immunoprophylaxis is based solely on Bacillus Calmette-Guérin (BCG) vaccination, and some of the new potential tuberculosis vaccines are based on the BCG genome. Therefore, it is reasonable to analyze the genomes of individual BCG substrains. The aim of this study was the genetic characterization of the BCG-Moreau Polish (PL) strain used for the production of the BCG vaccine in Poland since 1955. Sequencing of different BCG lots showed that the strain was stable over a period of 59 years. As a result of comparison, BCG-Moreau PL with BCG-Moreau Rio de Janeiro (RDJ) 143 single nucleotide polymorphisms (SNPs) and 32 insertion/deletion mutations (INDELs) were identified. However, the verification of these mutations showed that the most significant were accumulated in the BCG-Moreau RDJ genome. The mutations unique to the Polish strain genome are 1 SNP and 2 INDEL. The strategy of combining short-read sequencing with long-read sequencing is currently the most optimal approach for sequencing bacterial genomes. With this approach, the only available genomic sequence of BCG-Moreau PL was obtained. This sequence will primarily be a reference point in the genetic control of the stability of the vaccine strain in the future. The results enrich knowledge about the microevolution and attenuation of the BCG vaccine substrains.
The whole genome sequence obtained is the only genomic sequence of the strain that has been used for vaccine production in Poland since 1955. Sequencing of different BCG lots showed that the strain was stable over a period of 59 years. The comprehensive genomic analysis performed not only enriches knowledge about the microevolution and attenuation of the BCG vaccine substrains but also enables the utilization of identified markers as a reference point in the genetic control and identity tests of the stability of the vaccine strain in the future.
目前，结核病免疫预防仅基于卡介苗（BCG）接种，一些新的潜在结核病疫苗基于 BCG 基因组。因此，分析个体 BCG 亚株的基因组是合理的。本研究的目的是对波兰自 1955 年以来用于生产 BCG 疫苗的 BCG-Moreau Polish（PL）菌株的基因组进行遗传特征分析。对不同 BCG 批次的测序表明，该菌株在 59 年内保持稳定。通过比较，发现 BCG-Moreau PL 与 BCG-Moreau Rio de Janeiro（RDJ）143 个单核苷酸多态性（SNP）和 32 个插入/缺失突变（INDEL）。然而，对这些突变的验证表明，最显著的突变是在 BCG-Moreau RDJ 基因组中积累的。波兰菌株基因组特有的突变是 1 个 SNP 和 2 个 INDEL。将短读测序与长读测序相结合的策略是目前测序细菌基因组的最优化方法。通过这种方法，获得了 BCG-Moreau PL 唯一可用的基因组序列。该序列将主要成为未来疫苗菌株遗传稳定性控制的参考点。结果丰富了对 BCG 疫苗亚株微进化和衰减的认识。
获得的全基因组序列是自 1955 年以来波兰用于疫苗生产的唯一菌株基因组序列。对不同 BCG 批次的测序表明，该菌株在 59 年内保持稳定。进行的综合基因组分析不仅丰富了对 BCG 疫苗亚株微进化和衰减的认识，而且还使鉴定出的标记能够作为未来疫苗菌株遗传稳定性控制和身份测试的参考点加以利用。
