Characterization of gentamicin-resistant Mycoplasma hyorhinis.
A study was conducted to determine if a gentamicin-resistant strain of mycoplasma could be developed for use in validating current mycoplasma detection methods for biologic product harvest cell culture fluid (CCF) containing gentamicin. A strain of gentamicin-resistant Mycoplasma hyorhinis was isolated and characterized. The study showed that this organism was similar to the wild-type strain in all ways examined except gentamicin resistance. Both strains of mycoplasma (the gentamicin resistant and the wild-type) exhibited comparable growth patterns and showed 100% homology based on DNA sequencing and analysis of a 464-bp PCR product. Also, analysis using species-specific antisera identified both strains as M. hyorhinis. Two commonly used lot release mycoplasma detection methods (culture and DNAF) consistently detected mycoplasmas in spiked biologic product harvest CCF containing gentamicin but not in unspiked samples. This study demonstrates the first isolation and characterization of a gentamicin-resistant M. hyorhinis that can be used to validate mycoplasma detection methods for biologic product harvest CCF containing gentamicin.
Idowu
Arit D
AD
Quality Control Biology, Genentech, Inc., 73B, One, DNA Way, South San Francisco, CA 94080-4990, USA. arit@gene.com
Celones
Michelle C
MC
Salgado
Amparo E
AE
Marquez
Kristine M
KM
Pleibel
Nancy
N
D000818:Animals; D001431:Bacteriological Techniques; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D024881:Drug Resistance, Bacterial; D005839:Gentamicins / Q000494:pharmacology*; D006801:Humans; D045790:Mycoplasma hyorhinis / Q000187:drug effects* / Q000254:growth & development / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D003470:Culture Media; D004269:DNA, Bacterial; D005839:Gentamicins