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Clinical Laboratory Jun 2024The objective of this study is to understand the characteristics of the common spectrum of pathogen and the resistance of Mycoplasma in Sialidase-positive bacterial...
BACKGROUND
The objective of this study is to understand the characteristics of the common spectrum of pathogen and the resistance of Mycoplasma in Sialidase-positive bacterial vaginosis.
METHODS
The vaginal secretion specimens collected from August 2018 to October 2018 for the analysis of bacterial vaginosis (BV) were subjected to various techniques. These included routine leukorrhea examination, bacterial vaginosis sialidase testing, routine culture for common pathogens, mass spectrometry identification, and Mycoplasma resistance testing.
RESULTS
A total of 238 patients with BV were identified. The cleanliness grading was mostly clean (+) and clean (2+), accounting for 38.24% and 30.67%, respectively. The bacterial vaginosis test for vaginal secretions showed leukocyte esterase positivity in 220 cases, resulting in a positivity rate of 92.44%. The spectrum of routine culture was analyzed and divided into four groups: A, B, C, and D. Group A consisted of Candidal vaginitis (13.45%); group B consisted of Gardnerella vaginalis vaginitis (32.77%); group C consisted of gram-negative bacillus vaginitis (46.22%); and group D consisted of Streptococcus agalactiae vaginitis (7.56%). The identification and antimicrobial susceptibility testing results for Mycoplasma showed a high detection rate of BV, with a positivity rate of 86.13%. There was a high sensitivity to tetracyclines for Ureaplasma urealyticum and Mycoplasma hominis, but a high resistance to macrolides and quinolones.
CONCLUSIONS
Bacterial vaginosis existed in various complex forms, including Candida, Gardnerella vaginalis, Gram-negative bacillus, and Streptococcus agalactiae types. Moreover, there was an increasing trend of multi-drug resistance in Mycoplasma hominis. Therefore, it is crucial to pay attention to this condition and make accurate judgments based on the etiological characteristics and common antimicrobial susceptibility tests. This will enable the implementation of effective therapeutic interventions.
Topics: Humans; Female; Vaginosis, Bacterial; Neuraminidase; Mycoplasma; Adult; Drug Resistance, Bacterial; Vagina; Young Adult; Anti-Bacterial Agents; Mycoplasma Infections; Microbial Sensitivity Tests; Middle Aged; Adolescent
PubMed: 38868882
DOI: 10.7754/Clin.Lab.2024.230826 -
Journal of Clinical Microbiology Jun 2024Antimicrobial susceptibility testing (AST) of human mycoplasmas using microdilution is time-consuming. In this study, we compared the performance of MICRONAUT-S plates...
Evaluation of commercial, customized microdilution plates for , , and antimicrobial susceptibility testing and determination of antimicrobial resistance prevalence in France.
UNLABELLED
Antimicrobial susceptibility testing (AST) of human mycoplasmas using microdilution is time-consuming. In this study, we compared the performance of MICRONAUT-S plates (Biocentric-Bruker) designed for AST of , , and with the results using the Clinical & Laboratory Standards Institute (CLSI) reference method. Then, we investigated the prevalence and mechanisms of resistance to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. The two methods were compared using 60 strains. For the resistance prevalence study, -, -, and -positive clinical specimens were collected for 1 month each year in 22 French diagnostic laboratories. MICs were determined using the MICRONAUT-S plates. The (M) gene was screened using PCR, and fluoroquinolone resistance-associated mutations were screened using PCR and Sanger sequencing. Comparing the methods, 99.5% (679/680) MICs obtained using the MICRONAUT-S plates concurred with those obtained using the CLSI reference method. For 90 . isolates, the tetracycline, levofloxacin, and moxifloxacin resistance rates were 11.1%, 2.2%, and 2.2%, respectively, with no clindamycin resistance. For 248 . isolates, the levofloxacin and moxifloxacin resistance rates were 5.2% and 0.8%, respectively; they were 2.9% and 1.5% in 68 . isolates. Tetracycline resistance in (11.8%) was significantly ( < 0.001) higher than in (1.2%). No macrolide resistance was observed. Overall, the customized MICRONAUT-S plates are a reliable, convenient tool for AST of human mycoplasmas. Tetracycline and fluoroquinolone resistance remain limited in France. However, the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in spp. from 2010 to 2015 and requires monitoring.
IMPORTANCE
Antimicrobial susceptibility testing of human urogenital mycoplasmas using the CLSI reference broth microdilution method is time-consuming and requires the laborious preparation of antimicrobial stock solutions. Here, we validated the use of reliable, convenient plates designed for antimicrobial susceptibility testing that allows the simultaneous determination of the MICs of eight antibiotics of interest. We then investigated the prevalence and mechanisms of resistance of each of these bacteria to tetracyclines, fluoroquinolones, and macrolides in France in 2020 and 2021. We showed that the prevalence of levofloxacin and moxifloxacin resistance has increased significantly in spp. from 2010 to 2015 and requires ongoing monitoring.
PubMed: 38832769
DOI: 10.1128/jcm.00226-24 -
Biosensors May 2024Sexually transmitted diseases (STDs) are a global concern because approximately 1 million new cases emerge daily. Most STDs are curable, but if left untreated, they can...
Sexually transmitted diseases (STDs) are a global concern because approximately 1 million new cases emerge daily. Most STDs are curable, but if left untreated, they can cause severe long-term health implications, including infertility and even death. Therefore, a test enabling rapid and accurate screening and genotyping of STD pathogens is highly awaited. Herein, we present the development of the DNA-based 6STD Genotyping 9G Membrane test, a lateral flow strip membrane assay, for the detection and genotyping of six STD pathogens, including , , , , , and . Here, we developed a multiplex PCR primer set that allows PCR amplification of genomic materials for these six STD pathogens. We also developed the six ssDNA probes that allow highly efficient detection of the six STD pathogens. The 6STD Genotyping 9G Membrane test lets us obtain the final detection and genotyping results in less than 30 m after PCR at 25 °C. The accuracy of the 6STD Genotyping 9G membrane test in STD genotyping was confirmed by its 100% concordance with the sequencing results of 120 clinical samples. Therefore, the 6STD Genotyping 9G Membrane test emerges as a promising diagnostic tool for precise STD genotyping, facilitating informed decision-making in clinical practice.
Topics: Humans; Chlamydia trachomatis; Neisseria gonorrhoeae; Sexually Transmitted Diseases; Genotype; Trichomonas vaginalis; Genotyping Techniques; Mycoplasma hominis; Ureaplasma urealyticum; DNA; Mycoplasma genitalium; Biosensing Techniques; DNA, Bacterial; Multiplex Polymerase Chain Reaction
PubMed: 38785734
DOI: 10.3390/bios14050260 -
Frontiers in Cellular and Infection... 2024() belongs to the class , characterized by a very small genome size, reduction of metabolic pathways, including transcription factors, and the absence of a cell wall....
INTRODUCTION
() belongs to the class , characterized by a very small genome size, reduction of metabolic pathways, including transcription factors, and the absence of a cell wall. Despite this, they adapt well not only to specific niches within the host organism but can also spread throughout the body, colonizing various organs and tissues. The adaptation mechanisms of , as well as their regulatory pathways, are poorly understood. It is known that, when adapting to adverse conditions, can undergo phenotypic switches that may persist for several generations.
METHODS
To investigate the adaptive properties of related to survival in the host, we conducted a comparative phenotypic and proteogenomic analysis of eight clinical isolates of obtained from patients with urogenital infections and the laboratory strain H-34.
RESULTS
We have shown that clinical isolates differ in phenotypic features from the laboratory strain, form biofilms more effectively and show resistance to ofloxacin. The comparative proteogenomic analysis revealed that, unlike the laboratory strain, the clinical isolates possess several features related to stress survival: they switch carbon metabolism, activating the energetically least advantageous pathway of nucleoside utilization, which allows slowing down cellular processes and transitioning to a starvation state; they reconfigure the repertoire of membrane proteins; they have integrative conjugative elements in their genomes, which are key mediators of horizontal gene transfer. The upregulation of the methylating subunit of the restriction-modification (RM) system type I and the additional components of RM systems found in clinical isolates suggest that DNA methylation may play a role in regulating the adaptation mechanisms of in the host organism. It has been shown that based on the proteogenomic profile, namely the genome sequence, protein content, composition of the RM systems and additional subunits HsdM, HsdS and HsdR, composition and number of transposable elements, as well as the sequence of the main variable antigen Vaa, we can divide clinical isolates into two phenotypes: typical colonies (TC), which have a high growth rate, and atypical (aTC) mini-colonies, which have a slow growth rate and exhibit properties similar to persisters.
DISCUSSION
We believe that the key mechanism of adaptation of in the host is phenotypic restructuring, leading to a slowing down cellular processes and the formation of small atypical colonies. This is due to a switch in carbon metabolism and activation the pathway of nucleoside utilization. We hypothesize that DNA methylation may play a role in regulating this switch.
Topics: Humans; Mycoplasma hominis; Proteogenomics; Mycoplasma Infections; Adaptation, Physiological; Biofilms; Genome, Bacterial; Phenotype; Anti-Bacterial Agents; Bacterial Proteins; Drug Resistance, Bacterial
PubMed: 38756231
DOI: 10.3389/fcimb.2024.1398706 -
Journal of Thoracic Disease Apr 2024As a culture-independent method, metagenomic next-generation sequencing (mNGS) is widely used in microbiological diagnosis with advantages in identifying potential...
BACKGROUND
As a culture-independent method, metagenomic next-generation sequencing (mNGS) is widely used in microbiological diagnosis with advantages in identifying potential pathogens, guiding antibiotic therapy, and improving clinical prognosis, especially in culture-negative cases. () mediastinitis is a rare and severe disease for which etiological diagnosis is important but challenging. The application of mNGS in the etiological diagnosis of mediastinitis has seldom been studied.
METHODS
By searching the electronic medical history retrieval system with "" and "mediastinitis", seven patients diagnosed with mediastinitis were reviewed in Zhongshan Hospital, Fudan University, Shanghai from 9 December 2020 to 14 February 2023. Microbiological cultures and mNGS were conducted for blood, abscess, and/or mediastinal fluid. Adjustment of the antibiotic therapy due to mNGS was assessed. A literature review was conducted in the PubMed database beginning in 1970 for infection and mediastinitis.
RESULTS
For the seven patients, cultures of blood, abscess, and mediastinal fluid were negative whereas mNGS identified in serum, abscess, and/or mediastinal fluid and was used to guide specific antibiotic therapy. The stringent mapped reads number of genera (SMRNG), stringent mapped reads number of species (SMRN), and coverage rate of detection by mNGS were significantly higher in body fluid (abscess or mediastinal fluid) than in serum. All seven patients had underlying heart diseases and underwent previous cardiac surgery. The most common symptoms were fever and sternal pain. After detection of , antibiotics were adjusted to quinolones or doxycycline except for one patient, whose diagnosis was clarified after death. Two patients died. Literature review since 1970 identified 30 cases of extra-genital infection caused by . Including our seven new cases, 2 (5.4%) were neonates and 35 (94.6%) were adults. Thirty (81.1%) cases were postoperative infection and 15 (40.5%) had implanted devices. Five patients (13.5%) died.
CONCLUSIONS
mNGS might be a promising technology in the detection of fastidious pathogens such as . Accurate etiological diagnosis by mNGS could guide antibiotic therapy and facilitate clinical management.
PubMed: 38738251
DOI: 10.21037/jtd-24-286 -
Ginekologia Polska May 2024Genitourinary tract infections in pregnant women are one of the causes of abnormal pregnancy development including miscarriages, premature labor or premature rupture of...
OBJECTIVES
Genitourinary tract infections in pregnant women are one of the causes of abnormal pregnancy development including miscarriages, premature labor or premature rupture of membranes (PPROM). Atypical bacteria responsible for reproductive tract infections include Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum and Ureaplasma parvum. Identification of pathogens and appropriately selected therapy can improve obstetric outcomes in patients with symptoms of threatened miscarriage or threatened preterm labor. The purpose of our study is to analyze the impact of reproductive tract infections with ureaplasma and mycoplasma bacteria during pregnancy.
MATERIAL AND METHODS
In the presented study, we retrospectively analyzed the cases of 201 pregnant patients hospitalized in the Obstetrics and Gynecology Department of Poznan Regional Hospital in 2019-2022, who had a swab taken from external os area of the cervix for atypical bacteria - Ureaplasma and Mycoplasma. Only patients with symptoms of threatened miscarriage or threatened preterm labor were included in the study group. Microbiological tests were performed in the hospital laboratory with the Mycoplasma IST 3 test from Biomerieux.
RESULTS
We found a higher incidence of preterm labor in patients with symptoms of threatened preterm labor and a genital tract infection with Ureaplasma/Mycoplasma bacteria, compared to patients not infected with Mycoplasma/Ureaplasma - 31.1% vs 20% (p = 0.098). This observation in the case of Ureaplasma/Mycoplasma monoinfection group applied to 6 patients. - 75% of the group. Pregnant patients who had co-infection with other types of bacteria (48 patients in total) gave birth before 37 weeks of pregnancy in 27.1% of cases. We obtained a significant difference (p = 0.007) when comparing groups with positive and negative cultures for Ureaplasma/Mycoplasma by the presence of monoinfection/coinfection and the week of pregnancy in which delivery occurred. We also noted the effect of atypical bacterial infection for PPROM - this complication preceded preterm delivery in 40% of ureaplasma-positive patients, compared to 20% of PPROM without infection. We found a similar rate of preterm labor and pregnancy loss in Ureaplasma/Mycoplasma-positive patients who received antibiotic therapy (35.7%) compared to a group of pregnant women who did not receive treatment (31.6%).
CONCLUSIONS
Infection of the genital tract with atypical bacteria Ureaplasma and Mycoplasma has a negative impact on the course of pregnancy. Identification of the type of microorganisms in cervical canal secretions of pregnant patients with symptoms of threatened miscarriage or preterm labor seems crucial. The impact of antibiotic therapy though, requires further analysis.
PubMed: 38717222
DOI: 10.5603/gpl.99827 -
The New Microbiologica May 2024Trichomonas vaginalis and Mycoplasma hominis, two microorganisms causing infections of the urogenital tract, are closely associated in that they establish an...
The presence of Trichomonas vaginalis in urogenital samples can affect the sensitivity of Mycoplasma hominis identification techniques, leading to an underestimation of bacterial infections.
Trichomonas vaginalis and Mycoplasma hominis, two microorganisms causing infections of the urogenital tract, are closely associated in that they establish an endosymbiosis relationship, the only case among human pathogens. As a result, the presence of one microorganism may be considered a sign that the other is present as well. Identification of the two pathogens in clinical samples is based on cultivation techniques on specific media, even though in recent years, new sensitive and rapid molecular techniques have become. Here, we demonstrate that the concomitant presence of T.vaginalis in urogenital swabs may lead to a delay in the identification of M.hominis, and thus to an underestimation of bacterial infections when cultural techniques are used.
Topics: Mycoplasma hominis; Trichomonas vaginalis; Humans; Mycoplasma Infections; Female; Trichomonas Vaginitis; Male; Sensitivity and Specificity; Urogenital System; Adult
PubMed: 38700890
DOI: No ID Found -
Zhongguo Dang Dai Er Ke Za Zhi =... Apr 2024The patient, a male newborn, was admitted to the hospital 2 hours after birth due to prematurity (gestational age 27 weeks) and respiratory distress occurring 2 hours...
The patient, a male newborn, was admitted to the hospital 2 hours after birth due to prematurity (gestational age 27 weeks) and respiratory distress occurring 2 hours postnatally. After admission, the infant developed fever and elevated C-reactive protein levels. On the fourth day after birth, metagenomic next-generation sequencing of cerebrospinal fluid indicated a positive result for (9 898 reads). On the eighth day, a retest of cerebrospinal fluid metagenomics confirmed (56 806 reads). The diagnosis of purulent meningitis caused by was established, and the antibiotic treatment was switched to moxifloxacin [5 mg/(kg·day)] administered intravenously for a total of 4 weeks. After treatment, the patient's cerebrospinal fluid tests returned to normal, and he was discharged as cured on the 76th day after birth. This article focuses on the diagnosis and treatment of neonatal purulent meningitis, introducing the multidisciplinary diagnosis and treatment of the condition in extremely preterm infants.
Topics: Humans; Mycoplasma hominis; Infant, Newborn; Male; Infant, Extremely Premature; Moxifloxacin; Meningitis, Bacterial; Mycoplasma Infections; Anti-Bacterial Agents
PubMed: 38660910
DOI: 10.7499/j.issn.1008-8830.2312016 -
Frontiers in Cellular and Infection... 2024Bacterial vaginosis (BV) is a most common microbiological syndrome. The use of molecular methods, such as multiplex real-time PCR (mPCR) and next-generation sequencing,...
Vaginal microbiota molecular profiling and diagnostic performance of artificial intelligence-assisted multiplex PCR testing in women with bacterial vaginosis: a single-center experience.
BACKGROUND
Bacterial vaginosis (BV) is a most common microbiological syndrome. The use of molecular methods, such as multiplex real-time PCR (mPCR) and next-generation sequencing, has revolutionized our understanding of microbial communities. Here, we aimed to use a novel multiplex PCR test to evaluate the microbial composition and dominant lactobacilli in non-pregnant women with BV, and combined with machine learning algorithms to determine its diagnostic significance.
METHODS
Residual material of 288 samples of vaginal secretions derived from the vagina from healthy women and BV patients that were sent for routine diagnostics was collected and subjected to the mPCR test. Subsequently, Decision tree (DT), random forest (RF), and support vector machine (SVM) hybrid diagnostic models were constructed and validated in a cohort of 99 women that included 74 BV patients and 25 healthy controls, and a separate cohort of 189 women comprising 75 BV patients, 30 intermediate vaginal microbiota subjects and 84 healthy controls, respectively.
RESULTS
The rate or abundance of and were significantly reduced in BV-affected patients when compared with healthy women, while , , , BVAB2, 2, , and were significantly increased. Then the hybrid diagnostic models were constructed and validated by an independent cohort. The model constructed with support vector machine algorithm achieved excellent prediction performance (Area under curve: 0.969, sensitivity: 90.4%, specificity: 96.1%). Moreover, for subjects with a Nugent score of 4 to 6, the SVM-BV model might be more robust and sensitive than the Nugent scoring method.
CONCLUSION
The application of this mPCR test can be effectively used in key vaginal microbiota evaluation in women with BV, intermediate vaginal microbiota, and healthy women. In addition, this test may be used as an alternative to the clinical examination and Nugent scoring method in diagnosing BV.
Topics: Humans; Female; Vaginosis, Bacterial; Vagina; Adult; Microbiota; Multiplex Polymerase Chain Reaction; Young Adult; Artificial Intelligence; Lactobacillus; Support Vector Machine; Sensitivity and Specificity; ROC Curve; Middle Aged
PubMed: 38644962
DOI: 10.3389/fcimb.2024.1377225 -
Future Microbiology Apr 2024To study antimicrobial susceptibilities of genital mycoplasmas recovered from endocervical samples of reproductive-age, nonpregnant women ( = 8,336). For isolation...
To study antimicrobial susceptibilities of genital mycoplasmas recovered from endocervical samples of reproductive-age, nonpregnant women ( = 8,336). For isolation and susceptibility testing, the Mycoplasma IST2 kit was used. As many as 2093 samples were positive for mycoplasmas. The vast majority (>96%) of remained susceptible to tetracycline, doxycycline, josamycin and pristinamycin, whereas susceptibility rates to azithromycin and fluoroquinolones were significantly decreased. exhibited high susceptibility rates to doxycycline, pristinamycin and josamycin (98.1-100%), while susceptibilities to tetracycline and fluoroquinolones were considerably lower. Doxycycline remained highly potent for treating mycoplasmas; nevertheless, susceptibilities to other antimicrobials were significantly diminished.
PubMed: 38629933
DOI: 10.2217/fmb-2023-0238