VACCINE

Biologicals • November 2013
Detection of contaminants
in cell cultures, sera and trypsin
Author information
Pinheiro de Oliveira TF1, Fonseca AA Jr, Camargos MF,
de Oliveira AM, Pinto Cottorello AC, Souza Ados R, de Almeida IG, Heinemann MB.
Laboratório de Biologia Molecular/Laboratório de Diagnóstico de Doenças Virais
Laboratório Nacional Agropecuário de Minas Gerais
Pedro Leopoldo, Minas Gerais, Brazil
oliveiratfp@yahoo.com.br
Abstract
The aim of this study was standardization and application of polymerase chain reaction
(PCR) for the detection of contaminants in cell cultures, sera and trypsin. Five PCR protocols
were standardized to assess the presence of genetic material from mycoplasma,
porcine circovirus 1 (PCV1), bovine leukemia virus (BLV) or bovine viral diarrhea virus
(BVDV) in cell culture samples. PCR reactions for the genes GAPDH and beta-actin
were used to evaluate the efficiency of nucleic acid extraction. The PCR protocols were
applied to 88 cell culture samples from eight laboratories. The tests were also used to
assess potential contamination in 10 trypsin samples and 13 fetal calf serum samples
from different lots from five of the laboratories. The results showed the occurrence of the
following as DNA cell culture contaminants: 34.1% for mycoplasma, 35.2% for PCV1,
23.9% for BVDV RNA and 2.3% for BLV. In fetal calf sera and trypsin samples BVDV
RNA and PCV1 DNA was detected. The results demonstrated that cell culture, sera and
trypsin used by different laboratories show a high rate of contaminants. The results highlight
the need for monitoring cell cultures and controlling for biological contaminants in
laboratories and cell banks working with these materials.
“The results showed the
occurrence of the following
as DNA cell culture contaminants:
34.1% for mycoplasma,
35.2% for porcine circovirus 1,
23.9% for bovine viral diarrhea virus RNA
and 2.3% for bovine leukemia virus. The results
demonstrated that cell culture, sera and trypsin
used by different laboratories show a high rate
of contaminants. ”
http://www.ncbi.nlm.nih.gov/pubmed/?term=24071554