M. Tech. Veterinary Technology. / Ensuring that biological products (specifically vaccines) are free of bacterial
contamination presents a significant challenge. The European Pharmacopoeia
recommends the routine use of plates, broths and a filtration system for detection
of bacterial contaminants that might be present in raw materials. However, these
methods could not timeously detect slow growing bacterial contaminants and a
polymerase chain reaction was developed for the early detection of bacterial
contamination. To eliminate false positive results the researcher compared
ethidium monoazide bromide and propidium monoazide to distinguish between
viable and non-viable bacterial cells, using universal primers which amplified a
±1.5 kilobase pair fragment of 16S ribosomal ribonucleic acid. The results
indicated propidium monoazide more sensitive to detect low levels of viable
bacteria. A comparative test between European Pharmacopoeia recommended
and polymerase chain reaction methods found that the latter were the more
sensitive method for contamination detection. The sequencing results of positive
samples indicated the following contaminants: Stenotrophomonas spp. and
Pseudomonas spp. as pre-dominant followed by Bacillus spp., Enterobacter spp.,
Klebsiella spp. and Propionibacterium spp. Propidium monoazide combined with
polymerase chain reaction showed promise as a rapid, reproducible method for
accessing cell integrity and is valuable to those who wish to employ it as an early
detection method of possible contamination within a production system.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:tut/oai:encore.tut.ac.za:d1000706 |
| Date | January 2010 |
| Creators | Burger, Christina Elizabeth. |
| Contributors | Van Heerden, J. |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Text |
| Format | |
| Rights | © Tshwane University of Technology 2010 |
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