A Rapid Alternative to Culture Based Mycoplasma Detection

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S163-S164
Author(s):  
D. Bauer ◽  
J. Young ◽  
K. Norman
Keyword(s):  
Cells ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 1510 ◽  
Author(s):  
Anna Ligasová ◽  
Markéta Vydržalová ◽  
Renata Buriánová ◽  
Lenka Brůčková ◽  
Renata Večeřová ◽  
...  

Contamination of cell cultures by mycoplasmas is a very common phenomenon. As they can substantially alter cell metabolism and potentially spread to all cell cultures in laboratory, their early detection is necessary. One of the fastest and cheapest methods of mycoplasma detection relies on the direct staining of mycoplasmas’ DNA by DAPI or Hoechst dyes. Although this method is easy and fast to perform, it suffers from the low signal provided by these dyes compared to the nuclear DNA. Therefore, the reporter cell lines are used for cultivation of mycoplasmas before DAPI or the Hoechst staining step. In the study presented, we have developed and tested a new immunofluorescence assay for the detection of mycoplasmas. The method is based on the enzymatic labeling using DNA polymerase I and modified nucleotides utilizing nicks in the mycoplasmas’ DNA. Modified nucleotides are incorporated into mycoplasmas’ DNA and subsequently visualized by immunofluorescence microscopy. The developed approach is independent of the mycoplasma strain, does not intensely stain nuclear DNA, does not stain other bacteria, and provides higher sensitivity than the approach based on the direct labeling using DAPI or Hoechst dyes.


Biologicals ◽  
2013 ◽  
Vol 41 (6) ◽  
pp. 377-383 ◽  
Author(s):  
Alena Dabrazhynetskaya ◽  
Dmitriy V. Volokhov ◽  
Tsai-Lien Lin ◽  
Brian Beck ◽  
Rajesh K. Gupta ◽  
...  

Author(s):  
Ольга Островская ◽  
Olga Ostrovskaya ◽  
Марина Власова ◽  
Marina Vlasova ◽  
Ольга Кожарская ◽  
...  

The objective of the research was to study genital mycoplasma contribution to the development of reproductive disorders. Mycoplasma detection rate in genital smears of pregnant women of Khabarovsk was studied by a polymerase chain reaction (PCR) method. Ureaplasma (urealyticum+parvum) was detected in 56.3% of cases; Mycoplasma hominis in 7.5% of cases, Mycoplasma genitalium in 0% of cases. Genital mycoplasmas were detected in chorionic and fetal tissue samples in early-term spontaneous miscarriage cases, suction biopsies in women with recurrent miscarriage, tissue samples in elective abortions, placental samples of women whose pregnancy ended in preterm delivery, and placental samples of women who gave birth to children at term. Ureaplasma (urealyticum+parvum) DNA was detected in 11.9%, 18.8%, 1.4%, 32.6%, and 25.0% of cases, respectively; Mycoplasma hominis DNA in 7.4%, 12.5%, 2.6%, 2.3%, and 0% of cases, respectively; Mycoplasma genitalium DNA in 5.2%, 6.3%, 1.4%, 2.3%, 0%, respectively. Genital mycoplasmas were detected in autopsy material of children who had died in neonatal and postnatal periods. Relationship between mycoplasma concentration in vaginal flora of pregnant women and preterm delivery was identified using real-time PCR. Mycoplasma detection rate in reproductive pathologies was found to be significantly higher than in control groups. The results of the study favor the view of etiopathogenetic role of mycoplasmas in development of complications and outcomes of pregnancy.


2020 ◽  
Vol 276 ◽  
pp. 113769
Author(s):  
Brandy J. Russell ◽  
Kalanthe Horiuchi ◽  
Jason O. Velez ◽  
Christin H. Goodman ◽  
Barbara W. Johnson

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