Advances in PCR-based detection of mycoplasmas contaminating cell cultures.

Rawadi, Georges; Dussurget, Olivier

doi:10.1101/gr.4.4.199

Cited by 52 publications

(37 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Mycoplasmas, which belong to the class Mollicutes (Mycoplasma and Acholeplasma), are one of the most frequent contaminants found in cell cultures and also in virus stocks. 1 Among 100 Mycoplasma species that have been identiÞ ed, at least 20 Mycoplasma and Acholeplasma species have been found as cell culture/virus stock contaminants. 2,3 The frequency of Mycoplasma-infected cultures ranges from 10 to 50%.…”

Section: Membersmentioning

confidence: 99%

“…Polymerase chain reaction (PCR) based methods have been developed for detection of Mycoplasma and Acholeplasma contaminants in cell culture. 1,3,[8][9][10] These methods are relatively rapid, sensitive and can detect low copy number of Mycoplasmas present in cell cultures. Similar sensitive methods have also been used to know the possible aetiology of Mycoplasma infections in certain chronic diseases.…”

Section: Membersmentioning

confidence: 99%

See 1 more Smart Citation

Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Gopalkrishna

Verma

Kumbhar

et al. 2007

Indian J Med Microbiol

View full text Add to dashboard Cite

Section: Membersmentioning

confidence: 99%

Section: Membersmentioning

confidence: 99%

Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Gopalkrishna

Verma

Kumbhar

et al. 2007

Indian J Med Microbiol

View full text Add to dashboard Cite

“…One can distinguish two type of primer sets, mollicutes-specific primers that are able to detect all mycoplasmas contaminating cell cultures and species-specific primers that can be used to identify the species. This aspect was discussed elsewhere (Rawadi and Dussurget, 1995). Cross-reactivity of primers with other bacterial species and especially with the closely related clostridial species was reported by some investigators (Spaepen et al, 1992;Teyssou et al, 1993).…”

Section: Detection Of Mycoplasmas Contaminating Cell Culturesmentioning

confidence: 99%

“…It has been reported that samples may contain trace inhibitors of DNA polymerase enzymes producing false negative results_ Several techniques are available to set-up positive control_ Briefly, positive controls consist on the coamplification of a foreign plasmid with a distinct primers set or on the coamplification of a genetically modified DNA target with the same primers set. Recently, low stringency PCR has been used to detect mycoplasmas contaminating cell cultures and provides a reliable positive control (Rawadi and Dussurget, 1995). Laboratory practices necessary to minimize the hazard of contamination and inhibition prevented fast diffusion of PCR-based diagnosis, especially in clinical laboratories.…”

Section: Perspectivementioning

confidence: 99%

“…PCR has been widely and successfully applied to the detection of mycoplasmas contaminating cell cultures (van Kuppeveld etal., 1992(van Kuppeveld etal., , 1994Spaepen etal., 1992;Hopertet al, 1993;Rawadi et al, 1993;Teyssouet al, 1993;Roulland-Dussoix et al, 1994). Recent advances in this field have been reviewed elsewhere (Rawadi and Dussurget, 1995).…”

Section: Detection Of Mycoplasmas Contaminating Cell Culturesmentioning

confidence: 99%

See 1 more Smart Citation

Genotypic Methods for Diagnosis of Mycoplasmal Infections in Humans, Animals, Plants and Cell Cultures

Rawadi

Dussurget

1998

Biotechnology and Genetic Engineering Reviews

View full text Add to dashboard Cite

Establishment and characterization of 12 uterine cervical-carcinoma cell lines: Common sequence variation in theE7 gene of HPV-16-positive cell lines

Kim

Park

et al. 1997

Int. J. Cancer

View full text Add to dashboard Cite

Carcinoma of the uterine cervix is one of the most common neoplasms in women, and about 90% of all cervical carcinomas are epidemiologically associated with infection by high-risk human papillomaviruses (HPV) such as types 16, 18, 31 and 33 (Zur Hausen, 1991).Because most available cervical-cancer cell lines were initiated more than 20 years ago (Spence et al., 1988), they have been extensively passaged in culture for hundreds of times in uncontrolled growth conditions, and this might have resulted in changes in the original characteristics of the primary tumor (Spence et al., 1988;Braun et al., 1993). Moreover, because establishment is difficult, the numbers of recently established cell lines with a low passage number are limited (Spence et al., 1988;Braun et al., 1993). The establishment from histopathologically varied primary tumors of cervical-cancer cell lines with a low passage number and an understanding of their characteristics are therefore required.In cervical cancers, a high-risk viral DNA genome is often found to be integrated into the host chromosome by common disruption of the viral E1/E2 region (Schwarz et al., 1985;Chen et al., 1994). In in vitro experiments, E2 repress transcription from the promoters of the E6 and E7 genes. In contrast, E6/E7 genes are actively transcribed in E2-deleted cervical cancers and cell lines (Schwarz et al., 1985).E6 and E7 proteins have been shown to form complexes with the normal cellular protein, p53, or the retinoblastoma-susceptibility gene product, pRb, both of which are well-known tumor suppressors, and those complexes lead to inactivation of the suppression function . Oncogenic HPV-type E6 protein specifically binds to wild-type p53 protein and subsequently enhances the degradation of p53 protein in an ATP/ubiquitin-dependent manner (Scheffner et al., 1990). In addition, pRb is inactivated by binding with E7 protein or through mutation. E7 protein of high-risk HPVs complex with pRb to inactivate the tumor-suppressor function of pRb (reviewed by McIntyre et al., 1993). Functional relevance of alterations at the Rb-binding domain in the E7 gene remains unclear.In this context, we established 12 new uterine cervical-cancer cell lines (SNU-17, SNU-487, SNU-523, SNU-682, SNU-703, SNU-778, SNU-902, SNU-1000, SNU-1005, SNU-1160, SNU-1245 and SNU-1299) with early passages and described cell phenotypes, including histopathology of the primary tumor and in vitro growth characteristics. We also performed molecular characterization, including DNA-fingerprinting analysis and HPV detection, typing and determination of physical form (integrated or episomal) of viral DNA. RNA expression of HPV E6/E7 viral transcripts, sequence variation in the E7 gene of HPV and genetic alteration of p53 were also examined. MATERIAL AND METHODS Cell-line establishment and maintenanceCell lines were established from pathologically proven uterine cervical carcinomas (Table I). Solid tumors were finely minced with scissors and dissociated into small aggregates by pipetting. Appropriate amounts...

show abstract

Advances in PCR-based detection of mycoplasmas contaminating cell cultures.

Cited by 52 publications

References 52 publications

Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Detection of <i>Mycoplasma </i>species in cell culture by PCR and RFLP based method: Effect of BM-cyclin to cure infections

Genotypic Methods for Diagnosis of Mycoplasmal Infections in Humans, Animals, Plants and Cell Cultures

Establishment and characterization of 12 uterine cervical-carcinoma cell lines: Common sequence variation in theE7 gene of HPV-16-positive cell lines

Contact Info

Product

Resources

About