Optimized genotyping method for identification of bacterial contaminants in pharmaceutical industry.
Microbiological control is of crucial importance in the pharmaceutical industry regarding the possible bacterial contamination of the environment, water, raw materials and finished products. Molecular identification of bacterial contaminants based on DNA sequencing of the hypervariable 16SrRNA gene has been introduced recently. The aim of this study is to investigate the suitability of gene sequencing using our selection of PCR primers and conditions for rapid and accurate bacterial identification in pharmaceutical industry quality control. DNA was extracted from overnight incubated colonies from 10 bacterial ATCC strains, which are common contaminants in the pharmaceutical industry. A region of bacterial 16SrRNA gene was analyzed by bidirectional DNA sequencing. Bacterial identification based on partial sequencing of the 16SrRNA gene is the appropriate method that could be used in the pharmaceutical industry after adequate validations. We have successfully identified all tested bacteria with more than 99 % similarity to the already published sequences.
Borche
B
Stamatoski
Miroslava
M
Ilievska
Hristina
H
Babunovska
Nikola
N
Sekulovski
Sasho
S
Panov
D001419:Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D004340:Drug Contamination; D004345:Drug Industry; D005819:Genetic Markers; D005838:Genotype; D004364:Pharmaceutical Preparations / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D015203:Reproducibility of Results; D021521:Ribotyping / Q000379:methods*
D004269:DNA, Bacterial; D005819:Genetic Markers; D004364:Pharmaceutical Preparations; D012336:RNA, Ribosomal, 16S