Volume 5 • Issue 3 • 1000179 J Veterinar Sci Technolo ISSN: 2157-7579 JVST, an open access journal Open Access Research Article Veerasami et al., J Veterinar Sci Technolo 2014, 5:3 DOI: 10.4172/2157-7579.1000179 Keywords: Adventitious viruses; Multiplex PCR; Vaccines; Fetal bovine serum; Trypsin Introduction Tough infectious diseases are considered one of the major challenges for human and animal health across the world, the harmful efects of the pathogenic organisms have considerably lowered in recent times than a century ago. One of the signifcant factors contributing to this status is the availability of vaccines against infectious diseases coupled with vast improvements in vaccine technology and delivery systems. Eradication of smallpox from the entire world and the near elimination of wild polio virus is attributed to the availability of efective vaccines and its use. Similarly, a cattle plague, Rinderpest has been eradicated from the globe by systematic vaccination programs. Current research in vaccinology aims in developing safer and efcacious vaccines for control of infectious diseases. Vaccine safety is paramount because public fear of vaccination can dramatically reduce vaccine usage and thereby coverage. Tis would result in the subsequent reduction of herd immunity or in some cases even re-emergence of the disease. One of the emerging concerns on the vaccine safety is the possibility of contamination with adventitious viruses in vaccine batches [1]. Use of animal derived raw materials in vaccine development and manufacture is the major source for such contamination events [2,3]. Bovine serum and bovine/porcine trypsin are the most common animal derived raw materials in development and manufacturing of viral vaccines [4,5]. Identifcation of extraneous viruses or viral genomes, such as porcine circovirus and parvovirus, in rotavirus vaccine [6-9] and nodovirus in the insect cell derived Human papilloma virus vaccine, have created greater awareness among the regulatory agencies and vaccine industry about the need for extensive screening of vaccine banks, cell substrates and vaccine batches for adventitious viruses. Tere are several well- documented bovine and porcine viral contamination events such as Cachevalley virus [10], Reo viruses [11], Porcine circovirus [9], BVD [12], Pestivirus [13-16] and many other animal viruses [17] in live animal and human vaccines [18,19]. Currently, both the virus banks and cell seeds in vaccine *Corresponding author: Maroudam Veerasami, Translational Research Platform for Veterinary Biological, Tamil Nadu Veterinary and Animal Sciences University, India, Tel: 91-44-25556275, 25556276, 25556277, 25556278; E-mail: vmaroudam@tanuvas.org.in Received May 08, 2014; Accepted June 27, 2014; Published June 30, 2014 Citation: Veerasami M, Chitra K, Mohana Subramanian B, Thamaraikannan P, Srinivasan VA, et al. (2014) Individual and Multiplex PCR Assays for the Detection of Adventitious Bovine and Porcine Viral Genome Contaminants in the Commercial Vaccines and Animal Derived Raw Materials. J Veterinar Sci Technol 5: 179. doi:10.4172/2157-7579.1000179 Copyright: © 2014 Veerasami M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Animal derived raw materials such as trypsin and Fetal Bovine Serum are used in vaccine manufacturing and pose the threat of introducing animal pathogens as contaminants into the fnal products. Thus screening for adventitious virus/ genome is part of quality control in manufacturing of biologicals. Various in-vitro and in-vivo detection assays have been developed for the detection of potential viral contaminants in vaccines. However, these assays are expensive, time consuming, labor intensive and incomplete limiting their ability to meet the increasing demands of the biological industry. Polymerase chain reaction technology scores over the in-vitro and in-vivo assays in speed, specifcity, sensitivity and robustness of detection and can replace them in regular use. In the present study, a set of multiplex and individual PCRs were developed for the detection of porcine (n=6) and bovine viral genomes (n=5). Veterinary vaccines (10), human vaccines (9), porcine typsin lots (9), and fetal bovine serum (8) were screened for adventitious viral genomes using multiplex PCRs. It was observed that 60% of veterinary vaccines, 77.7% of trypsin, and 62.5% of fetal bovine serum were contaminated with adventitious viral genomes. Individual and Multiplex PCR Assays for the Detection of Adventitious Bovine and Porcine Viral Genome Contaminants in the Commercial Vaccines and Animal Derived Raw Materials Maroudam Veerasami 1 *, Chitra K 1 , Mohana Subramanian B 1 , Thamaraikannan P 1 , Srinivasan VA 2 and Dhinakar Raj G 1 1 Translational Research Platform for Veterinary Biological, Tamil Nadu Veterinary and Animal Sciences University, India 2 National Dairy Development Board, India manufacturing are tested for bovine and porcine adventitious agents in compliance with the regulations specifed in the 9CFR 113 by U.S. Department of Agriculture (USDA) for veterinary products [17]. However, the 9CFR procedures are laborious, expensive and time- consuming and incorporating this method for the testing of regular vaccine batches is difcult. Moreover, the 9CFR procedure does not address the exhaustive list of viruses that are potential contaminants in animal derived raw materials [20,21]. Te 9CFR procedure was essentially designed to detect the viruses that have the potential to infect bovine and porcine hosts. Since this procedure is also used to validate cell and virus banks of human vaccines, there is a possibility of overlooking the presence of zoonotic viruses [21]. Therefore, the need for additional testing procedures may be useful adjuncts to 9 CFR tests in quality control laboratories of vaccine manufacturers [3,19]. In recent times PCR assays have found wide application in genome detection in the diagnostics laboratories due to its speed, sensitivity, specificity and ease of use [22]. The present study describes a set of optimized multiplex and individual PCRs for detecting adventitious viral genome of porcine and bovine origin in the commercially available vaccines and animal derived raw materials. J o u r n a l o f V e t e r i n a r y S c i e n c e & T e c h n o l o g y ISSN: 2157-7579 Journal of Veterinary Science & Technology