RAJENDRAN VIJAYAKUMAR¹*, FAIZ ALFAIZ¹, TIM SANDLE²
*Corresponding author
1. Department of Biology, College of Science Zulfi, Majmaah University, Majmaah, Saudi Arabia
2. Bio Products Laboratory, Elstree, United Kingdom

Abstract

Rapid detection of microbial contaminants in pharmaceutical products is play a vital role in contamination control. The purpose of this study is to standardize multiplex polymerase chain reaction (mPCR) assay to detect the pharmaceutical contaminants (either bacterial or fungal) and Pseudomonas aeruginosa specifically in single PCR reaction. A total of 13 pharmaceutical samples of ophthalmic products, chemotherapeutic, psychiatric, cardiac and gastrointestinal drugs were selected and artificially contaminated with <10 CFU of microorganisms. After enrichment, DNA was extracted, targeting the conserved region in eubacterial genome (16SrRNA), panfungal genome (28S rRNA) and P. aeruginosa (oprL). Specificity and sensitivity of the technique were verified. The mPCR assay was standardized by varying concentration of PCR reagents and conditions. In conclusion, there was no PCR inhibition and false positive results were observed. This assay can easily be incorporated for pharmaceutical products to quickly screen for microbial contamination.

INTRODUCTION

Microbial contamination control is a major challenge for the pharmaceutical industry and regulatory agencies, as reflected in product recall data. As an example, and based on FDA enforcement reports from 2002-2019, recalls due to bacteria (including relatively high incidences of Burkholderia cepacia complex organisms (formerly Pseudomonas cepacia), fungi, together with findings relating to lack of sterility assurance, were higher compared with earlier review periods (1).  As well as B. cepacia being indicated as an organism of concern, among the sterile products contaminated with fungi, 95% were recalled due to Aspergillus contamination. Lack of sterility assurance was the most common reason for sterile drugs recalls during the period. The classes of drugs recalled ranged from injectable saline solutions, hormones, ophthalmic solutions, water for injection, antibiotics, vitamins ... ...