World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA

Nübling, C. Micha; Baylis, Sally A.; Hanschmann, Kay-Martin; Montag-Lessing, Thomas; Chudy, M.; Kreß, Julia; Ulrych, Ursula; Czurda, Stefan; Rosengarten, Renate

doi:10.1128/aem.01150-15

Cited by 30 publications

(17 citation statements)

References 16 publications

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“…Great variation was seen in assay performance at the lower concentrations. In addition to the QCMD program, an international standard for use with NAATs for detection of M. pneumoniae has been developed by the World Health Organization (337). Results of comparative studies clearly demonstrate the need for external proficiency testing to document accuracy of molecular testing for M. pneumoniae and the need for improvement of analytical sensitivity for some assays, as well as standardization of sample preparation, including DNA extraction, to achieve optimum results.…”

Section: Nucleic Acid Amplificationmentioning

confidence: 99%

Mycoplasma pneumoniae from the Respiratory Tract and Beyond

et al. 2017

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is an important cause of respiratory tract infections in children as well as adults that can range in severity from mild to life-threatening. Over the past several years there has been much new information published concerning infections caused by this organism. New molecular-based tests for detection are now commercially available in the United States, and advances in molecular typing systems have enhanced understanding of the epidemiology of infections. More strains have had their entire genome sequences published, providing additional insights into pathogenic mechanisms. Clinically significant acquired macrolide resistance has emerged worldwide and is now complicating treatment. susceptibility testing methods have been standardized, and several new drugs that may be effective against this organism are undergoing development. This review focuses on the many new developments that have occurred over the past several years that enhance our understanding of this microbe, which is among the smallest bacterial pathogens but one of great clinical importance.

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Section: Nucleic Acid Amplificationmentioning

confidence: 99%

Mycoplasma pneumoniae from the Respiratory Tract and Beyond

et al. 2017

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“…The contamination of cell cultures by mycoplasma is recognized as one of the most serious and persistent problems in cell culture, what culminates in a large source of false and non-reproducible scientific results (Armstrong et al, 2010;Laborde et al, 2010;Volokhov et al, 2011). Different cells may present diverse morphological and physiological signals when infected by mycoplasma (Nübling et al, 2015). HEp-2 cells are used in a wide range of biological assays and may be contaminated by mycoplasma.…”

Section: Discussionmentioning

confidence: 99%

“…Most of mycoplasma are extracellular parasites, with ability to adhere firmly to the membrane of the host cell. That-may cause changes in cell physiology and viability, growth rate, metabolism and gene expression (Nübling et al, 2015). The sources of contamination by mycoplasma are diverse: cross contamination from another cell culture, laboratory equipment, contaminated reagents, fetal bovine serum, cryogenic flasks, culture medium and experimental animals (Corral-Vázquez et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

“…They easily spread from one culture to another. The impact of mycoplasma contamination on cultures depend on the specie, the intensity of bacterial replication as well as the infected cell type (Nübling et al, 2015). Generally, there is a decline in the nutritional quality perceived of the culture medium (depletion of sugars, amino acids and fatty acids and cholesterols) (Masukagami et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

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Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma

et al. 2021

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Contamination of primary and cell cultures by mycoplasmas is one of the main economic and biological pitfalls in basic research, diagnosis and manufacture of biotechnological products. It is a common issue which may be difficult to conduct surveillance on. Mycoplasma presence may affect several physiological parameters of the cell, besides being considered an important source of inaccurate and/or non-reproducible scientific results. Each cell type presents characteristical symptoms, mainly morphological, that indicate a contamination by mycoplasma. HEp-2 cells originate from carcinoma of the larynx and are, therefore, part of the respiratory tract, which is one of mycoplasma habitats. Despite the importance these cells in several biological research (evaluation of cell proliferation and migration, apoptosis, antiviral and antitumor compounds), the alterations induced by mycoplasma contamination in HEp-2 cells have not yet been described. Here, we describe the progressive morphological alterations in culture of HEp-2 cells infected with mycoplasma, as well as the-diagnosis of the infection and its treatment. Mycoplasma contamination described within this work led to cytoplasm elongation, cell-to-cell spacing, thin plasma membrane projections, cytoplasmic vacuoles, fusion with neighboring cells, and, finally, cell death. Contamination was detected by fluorescence imaging (DAPI) and PCR reactions. The cultures were treated with BM-Cyclin antibiotic to eliminate contamination. The data presented here will be of relevance to researchers whose investigations involve cell culture, especially respiratory and HEp-2 cells.

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“…Therefore, it is paramount to evaluate the potential risk of passing cultures between different facilities and to institute a routine screening process (Young et al, 2010). Standardized screening practices for mycoplasma detection are preferred as the range of different available assays produces variable results and conclusions (Nubling et al, 2015).…”

Section: Mycoplasma Contaminationmentioning

confidence: 99%

Good Cell Culture Practice for stem cells and stem-cell-derived models

Pamies

2017

Toxicology Letters

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SummaryThe first guidance on Good Cell Culture Practice (GCCP) dates back to 2005. This document expands this to include aspects of quality assurance for in vitro cell culture focusing on the increasingly diverse cell types and culture formats used in research, product development, testing and manufacture of biotechnology products and cell-based medicines. It provides a set of basic principles of best practice that can be used in training new personnel, reviewing and improving local procedures, and helping to assure standard practices and conditions for the comparison of data between laboratories and experimentation performed at different times. This includes recommendations for the documentation and reporting of culture conditions. It is intended as guidance to facilitate the generation of reliable data from cell culture systems, and is not intended to conflict with local or higher level legislation or regulatory requirements. It may not be possible to meet all recommendations in this guidance for practical, legal or other reasons. However, when it is necessary to divert from the principles of GCCP, the risk of decreasing the quality of work and the safety of laboratory staff should be addressed and any conclusions or alternative approaches justified. This workshop report is considered a first step toward a revised GCCP 2.0.

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World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA

Cited by 30 publications

References 16 publications

Mycoplasma pneumoniae from the Respiratory Tract and Beyond

Mycoplasma pneumoniae from the Respiratory Tract and Beyond

Diagnosis and treatment of HEp-2 cells contaminated with mycoplasma

Good Cell Culture Practice for stem cells and stem-cell-derived models

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