Mycoplasma spp. contamination is a major concern in laboratories handling cell cultures, and routine detection methods are usually time-consuming, laborious and lack sensitivity. This study presents a streamlined workflow integrating rapid thermal DNA extraction (99 °C-1 min) with a SYBR Green-based qPCR for Mycoplasma detection. High-coverage primers targeting an 86-bp region of the 16S rDNA were designed using 109 Mycoplasma spp. sequences from GeneBank. In silico analysis confirmed full primer annealing to major cell culture contaminants (M. arginini, M. hominis, M. orale, and M. hyorhinis). Upon thermal lysis and qPCR optimization, the yield of the protocol was equivalent to that of phenol-chloroform extraction plus qPCR, with a detection limit of 64 bacterial cells. Finally, the performance of the protocol was confirmed in cell cultures with known Mycoplasma spp. contamination, accurately reproducing the contamination status. Thus, the developed protocol provides a simple, rapid, cost-effective, and sensitive method for monitoring Mycoplasma spp. in cell cultures.
- MeSH
- Cell Culture Techniques * methods MeSH
- DNA, Bacterial genetics isolation & purification MeSH
- DNA Primers genetics MeSH
- Real-Time Polymerase Chain Reaction * methods MeSH
- Humans MeSH
- Mycoplasma * genetics isolation & purification classification MeSH
- Workflow MeSH
- DNA, Ribosomal genetics MeSH
- RNA, Ribosomal, 16S genetics MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
OBJECTIVES: The aim of this study is to determine the incidence of Mycoplasma pneumoniae pneumonia (MPP) in children and adolescents in Prague, Czech Republic, between January and July 2024, and to compare the findings with data from the preceding period. METHODS: A retrospective analysis of data of paediatric patients at our single tertiary care facility was conducted. Two distinct patient cohorts were subjected to analysis: the first comprising individuals who had been hospitalised between January 2019 and July 2024, and the second consisting of outpatients who had been treated during the periods of January to July 2023 and January to July 2024. RESULTS: A 12.3-fold increase in the number of outpatients diagnosed with MPP was observed between January and July 2024 in comparison to the same period in 2023, with 111 cases reported in 2024 versus 9 cases in 2023. A total of 23 patients were hospitalised with MPP between January 2019 and July 2024, with 15 of these hospitalisations having occurred between January and July 2024. The median age was 12 years, with an age range of 1 to 17 years. The majority of cases presented with a high fever, chest pain, and required oxygen support. A failure of the clarithromycin treatment was observed, resulting in 19.48% of doxycycline prescriptions being issued due to a prior failure of clarithromycin treatment. During the monitoring period, no cases of treatment failure with doxycycline were documented. CONCLUSION: The present study demonstrates an emerging trend of increased incidence of Mycoplasma pneumoniae pneumonia in the paediatric population during the initial seven months of 2024 in the Czech Republic. Doxycycline has been demonstrated to be the optimal antibiotic for the treatment of MPP and in accordance with the prevailing practice in other states it should be included in the therapeutic regimen even in children under the age of eight. The authors put forward recommendations for the implementation of measures aimed at reducing the negative impact of MPP on public health.
- MeSH
- Anti-Bacterial Agents * therapeutic use MeSH
- Tertiary Care Centers MeSH
- Child MeSH
- Hospitalization statistics & numerical data MeSH
- Incidence MeSH
- Infant MeSH
- Humans MeSH
- Adolescent MeSH
- Mycoplasma pneumoniae MeSH
- Pneumonia, Mycoplasma * epidemiology drug therapy MeSH
- Child, Preschool MeSH
- Retrospective Studies MeSH
- Check Tag
- Child MeSH
- Infant MeSH
- Humans MeSH
- Adolescent MeSH
- Male MeSH
- Child, Preschool MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Geographicals
- Czech Republic MeSH
Following the COVID-19 infection, the sternum dislocation and wound dehiscence resulted in an infection complicating the recovery of an immunosuppressed patient after bilateral lung transplantation. Anaerobic culture (96 h) of milky cloudy wound secretion resulted in the growth of pinpoint haemolytic colonies identified as Metamycoplasma hominis (formerly Mycoplasma hominis). The search for the endogenous source of the infection found the bacterium exclusively in the patient's sputum, making a possible link to donor lung M. hominis colonization. Unfortunately, the donor samples were no longer available. The wound infection was successfully treated with 17 days of clindamycin despite the continuous PCR detection of M. hominis in the sputum after the end of the treatment.
- MeSH
- Anti-Bacterial Agents therapeutic use MeSH
- COVID-19 diagnosis MeSH
- Immunocompromised Host MeSH
- Surgical Wound Infection * microbiology drug therapy diagnosis MeSH
- Clindamycin therapeutic use MeSH
- Middle Aged MeSH
- Humans MeSH
- Mycoplasma hominis * genetics isolation & purification MeSH
- Mycoplasma Infections * microbiology diagnosis drug therapy MeSH
- SARS-CoV-2 genetics isolation & purification MeSH
- Sputum microbiology MeSH
- Lung Transplantation * adverse effects MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
- MeSH
- Anti-Bacterial Agents * therapeutic use MeSH
- Drug Resistance, Bacterial drug effects MeSH
- Microorganisms, Genetically-Modified MeSH
- Respiratory Tract Infections * drug therapy MeSH
- Clinical Studies as Topic MeSH
- Humans MeSH
- Mycoplasma pneumoniae isolation & purification MeSH
- Pseudomonas aeruginosa drug effects MeSH
- Check Tag
- Humans MeSH
- Publication type
- Newspaper Article MeSH
A study targeting the etiology of severe anaemia that sporadically occurred in laboratory-bred cyprinid hybrids resulted in a diagnosis of a Mycoplasma-like organism selectively invading the cytoplasm of erythrocytes. Despite the fact that there was a concurrent yeast infection in moribund anaemic hybrids, the primary role in the development of anaemia was assigned to the Mycoplasma-like organism due to its regular occurrence in erythrocytes of both the moribund hybrids and hybrids that were free of yeast infection yet showed early to advanced symptoms of the disease. Novel data on the Mycoplasma-like organism's cytoskeleton were obtained from ultrathin sections of affected erythrocytes. An ultrastructural study of the concurrent yeast infection in moribund hybrids manifesting the most advanced anaemia revealed the presence of Titan cells in ascitic fluid. The original findings presented in this study underline the diagnostic relevance of transmission electron microscopy in the research of similar infections.
- MeSH
- Anemia * veterinary MeSH
- Erythrocytes ultrastructure MeSH
- Mycoplasma * ultrastructure MeSH
- Mycoplasma Infections * diagnosis veterinary MeSH
- Saccharomyces cerevisiae MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- MeSH
- Ampicillin therapeutic use MeSH
- Anti-Bacterial Agents pharmacology therapeutic use MeSH
- Cefuroxime therapeutic use MeSH
- Child MeSH
- Clindamycin therapeutic use MeSH
- Humans MeSH
- Mycoplasma pneumoniae pathogenicity MeSH
- Pneumonia, Mycoplasma * diagnosis etiology drug therapy complications MeSH
- Pleural Effusion etiology MeSH
- Sulbactam therapeutic use MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Publication type
- Case Reports MeSH
Spektrum sexuálně přenosných infekcí, vyskytujících se v ČR, je velmi pestré a v zásadě kopíruje výskyt ve světě. V ČR jsou v současné době běžně diagnostikovány syfilis, kapavka, infekce Chlamydia trachomatis, Mycoplasma genitalium. Moderní diagnostické možnosti (polymerázová řetězová reakce, PCR) přinášejí velkou diagnostickou spolehlivost i uživatelský komfort. Největším problémem se jeví zvyšující se výskyt rezistence k antibiotikům a časté vícečetné koinfekce.
There is a really broad range of sexually transmitted infections presented in the Czech Republic. It is very close to a worldwide situation. Nowadays we can diagnose syphilis, gonorrhoea, Chlamydia trachomatis and Mycoplasma genitalium infections. Modern diagnostic approaches (polymerase chain reaction, PCR) bring reliability and user friendly format. The biggest problems are antimicrobial resistance and multiply co-infections.
- MeSH
- Drug Resistance, Microbial MeSH
- Chlamydia Infections diagnosis epidemiology drug therapy transmission MeSH
- Diagnosis, Differential MeSH
- Humans MeSH
- Lymphogranuloma Venereum diagnosis epidemiology drug therapy MeSH
- Mycoplasma genitalium pathogenicity drug effects MeSH
- Mycoplasma Infections diagnosis epidemiology drug therapy transmission MeSH
- Neisseria gonorrhoeae pathogenicity drug effects MeSH
- Polymerase Chain Reaction methods MeSH
- Sexually Transmitted Diseases * epidemiology drug therapy microbiology MeSH
- Syphilis diagnosis drug therapy classification MeSH
- Treponema pallidum pathogenicity drug effects MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Review MeSH
Ve druhém pokračování moderní diagnostiky sexuálně přenosných infekcí se budeme věnovat novým a méně obvyklým bakteriálním původcům onemocnění. S ohledem na běžný výskyt koinfekcí v této oblasti je důležité využití multiplexního diagnostického přístupu - PCR. Šířeji vedená diagnostika zejména u rizikových pacientů zkracuje dobu antibiotické léčby a riziko klinických relapsů. Vždy je vhodné vyloučit i infekci HIV, pravděpodobnost přenosu je několikanásobně vyšší.
The second part of modern diagnosing of sexually transmitted infections deals with new and less common bacterial causes of the diseases. Given the frequency of coinfections in this setting, it is important to use a multiplex diagnostic approach - PCR. A broader diagnostic workup, particularly in patients at risk, reduces the duration of antibiotic therapy and the risk of clinical relapses. It is always advisable to rule out HIV infection; the likelihood of transmission is several-fold higher.
- MeSH
- Granuloma Inguinale diagnosis microbiology therapy MeSH
- Haemophilus ducreyi genetics isolation & purification pathogenicity MeSH
- Humans MeSH
- Chancroid diagnosis microbiology therapy MeSH
- Mycoplasma genetics isolation & purification pathogenicity MeSH
- Sexually Transmitted Diseases * diagnosis therapy MeSH
- Check Tag
- Humans MeSH
- Publication type
- Review MeSH
BACKGROUND: To determine the association between microbial invasion of the amniotic cavity (MIAC) and the presence of Lactobacillus crispatus- or Lactobacillus iners-dominated cervical microbiota in pregnancies with preterm prelabor rupture of membrane. Next, to assess the relationship between the presence of L. crispatus- or L. iners-dominated cervical microbiota and short-term neonatal morbidity. METHOD: A total of 311 women were included. Cervical samples were obtained using a Dacron polyester swab and amniotic fluid samples were obtained by transabdominal amniocentesis. Bacterial DNA, L. crispatus, and L. iners in the cervical samples were assessed by PCR. Cervical microbiota was assigned as L. crispatus- or L. iners-dominated when the relative abundance of L. crispatus or L. iners was ≥50% of the whole cervical microbiota, respectively. RESULTS: Women with MIAC showed a lower rate of L. crispatus-dominated cervical microbiota (21% vs. 39%; p = 0.003) than those without MIAC. Lactobacillus crispatus-dominated cervical microbiota was associated with a lower rate of early-onset sepsis (0% vs. 5%; p = 0.02). CONCLUSIONS: The presence of L. crispatus-dominated cervical microbiota in women with preterm prelabor rupture of membrane was associated with a lower risk of intra-amniotic complications and subsequent development of early-onset sepsis of newborns.
- MeSH
- Amniocentesis methods MeSH
- Cervix Uteri microbiology MeSH
- Chlamydia trachomatis MeSH
- Chorioamnionitis microbiology MeSH
- Lactobacillus crispatus * MeSH
- Lactobacillus * MeSH
- Humans MeSH
- Microbiota MeSH
- Mycoplasma hominis MeSH
- Infant, Newborn MeSH
- Amniotic Fluid microbiology MeSH
- Obstetric Labor, Premature MeSH
- Fetal Membranes, Premature Rupture microbiology MeSH
- Retrospective Studies MeSH
- Pregnancy MeSH
- Ureaplasma MeSH
- Check Tag
- Humans MeSH
- Infant, Newborn MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
