scholarly journals Laboratory diagnostics of urogenital mycoplasma colonization and infection

2021 ◽  
Vol 51 (4) ◽  
pp. 68-72
Author(s):  
L. N. Novikova ◽  
A. E. Taraskina

The paper presents data on the possibilities of determining urogenital mycoplasmas (Mycoplasma hominis and Ureaplasma urealyticum) using some variants of culture and PCR methods. The scheme of cultural studies in full is given, with the help of which high specificity and sensitivity indicators are achieved. The proposed research algorithm for detecting Mycoplasma hominis and Ureaplasma urealyticum, based on the use of culture and PCR methods as complementary.

Author(s):  
Zhaoyang Sun ◽  
Jun Meng ◽  
Su Wang ◽  
Feng Yang ◽  
Tao liu ◽  
...  

BackgroundSexually transmitted infections (STIs) are some of the most common communicable conditions and exert impact on the health and lives of many hundreds of millions of people across the world every year. Screening high-risk populations and conducting comprehensive detection tests would lead to a significant improvement in preventing the transmission of STIs and help us to provide rapid treatment to those affected. Here, we successfully established and validated a novel high-throughput multiplex gene detection system (HMGS) for the simultaneous and semiquantitative detection of six important curable sexually transmitted pathogens in a single reaction from secretions samples.MethodFluorescently labeled primers were designed to target specific conserved and single-copy gene fragments of Ureaplasma urealyticum (U. urealyticum), Mycoplasma hominis (M. hominis), Chlamydia trachomatis (C. trachomatis), Neisseria gonorrhoeae (N. gonorrhoeae), Trichomonas vaginalis (T. vaginalis), and Treponema pallidum (T. pallidum). The specificity and sensitivity of the STI-HMGS was validated and optimized using plasmids and quantitative genomic DNA. Next, we validated the performances of the STI-HMGS for clinical application by testing samples of clinical secretions collected from patients who visited the gynecology and urology outpatient clinics of our reproductive medicine center. Results derived from the STI-HMGS were then compared with three approved commercialized kits that used to detect U. urealyticum, C. trachomatis and N. gonorrhoeae, respectively, followed by further validation with Sanger sequencing for all pathogens. Finally, a comprehensive analysis of epidemiology was performed among different subgroups to investigate the association between infection rates and clinically-relevant information.ResultsThe sensitivity of STI-HMGS for six target genes was 10 copies/µL. Data derived from the detection of 381 clinical secretions demonstrated that the STI-HMGS exhibited high concordance rate compared with approved commercialized kits and almost 100% sensitivity and specificity for the detection of six sexually transmitted pathogens when validated by Sanger sequencing. Semi-quantitative analysis found that STIs caused by N. gonorrhoeae had a significantly higher (P<0.05) pathogen load than the other pathogens. Infections caused by C. trachomatis were significantly more common in younger individuals (P<0.05). We also found that U. urealyticum infections were more likely to happen in females; while the males were more affected by N. gonorrhoeae (P<0.05).ConclusionsSTI-HMGS proved to be an efficient method for the semi-quantitative detection of six important curable sexually transmitted pathogens and therefore represents an alternative method for the clinical detection and monitoring of STIs.


2014 ◽  
Vol 5 (1) ◽  
pp. 45-48
Author(s):  
K. V. Bubalo ◽  
L. P. Golodok ◽  
A. I. Vinnikov

The frequency of urogenital mycoplasmas detection in women of different ages was studied in culture with the help of DUO test-system in order to determine their etiological significance in the development of inflammatory processes of women urogenital tract. We identified the researched cultures Mycoplasma hominis, Ureaplasma urealyticum in the diagnostic titer >104 TEM/ml indicating severe contamination by microorganisms, and in the titer <103 TEM/ml, the carrier state of the identified microorganisms. Of 120 studied isolates of women urogenital tract there have been identified 113 strains of genital mycoplasmas, among which 63%  – U. urealyticum, 32% – M. hominis, 3% – microbial association of U. urealyticum – M. hominis. According to the study of frequency of detection of urogenital mycoplasma using DUO test-system culture method, it was found that the most frequently observed ones were U. urealyticum in 75 women (63%) of all individuals, M. hominis in 38 women (32%) in different diagnostic titers (>104 TEM/ml, <103 TEM/ml) in 4 women (3%) U. urealyticum – M. hominis was observed in microbial associations and mycoplasma were not found in 3 women (2%) of all surveyed patients. U. urealyticum and M. hominis in the diagnostic titer of >104 TEM/ml was observed in 55 women (46%) and 20 women (17%), respectively, and the titer of <103 CFU/ml U. urealyticum was observed in 20 women (17%), and M. hominis in 18 women (15%). Analysis of genital mycoplasmas distribution among women of different ages has shown that there was the certain correlation between the patient age and frequency of genital mycoplasmas detection: the highest detection rate was observed in women age of 24–29. The dominant pathogen of urogenital tract inflammatory processes in women in 24–29 age group is U. urealyticum. The comparison of DUO test-system and PCR data has shown that DUO test-system in culture allowed more sensitive quantitave characterization of mycoplasmas, however, for the more effective laboratory diagnostics it was necessary to use complex methods to increase the probability of pathogen detection. Incidence of mycoplasmas in women with the presence of inflammation was higher than in women having the inflammation in the genital tract. In this case, potential symptom-free carriers exist for the development of inflammation of urogenital tract of women. Scientists have proved that mycoplasma could cause vulvovaginitis, urethritis, paraurethritis, bartholinitis, adnexitis, salpingitis, endometritis, and ovaritis. 


2003 ◽  
Vol 57 (1-2) ◽  
pp. 63-72
Author(s):  
Slavko Djurisic ◽  
Sava Lazic ◽  
Tamas Petrovic ◽  
Sara Savic-Jevdjenic ◽  
Diana Lupulovic

The ELISA technique is one of the most commonly used laboratory-diagnostical methods for diagnosing many infective diseases in humans and animals today. Due to its high specificity and sensitivity this technique is considered a method of choice in diagnosing many viral diseases. The phases and components of the ELISA technique are constantly improved because of its mass use, as well as the high demands of laboratory diagnostics. Therefore it is necessary to continually inform the expert public of all the innovations, forms and possibilities of the ELISA technique. This paper describes the factors that caused the genesis of the ELISA technique and shows its forms. The methodological and practical aspects of the ELISA technique, when applied in laboratory diagnostics of veterinary medicine are also presented.


2020 ◽  
Vol 20 (10) ◽  
pp. 831-840
Author(s):  
Weibin Li

Sepsis is still a severe health problem worldwide with high morbidity and mortality. Blood bacterial culture remains the gold standard for the detection of pathogenic bacteria in bloodstream infections, but it is time-consuming, and both the sophisticated equipment and well-trained personnel are required. Immunoassays and genetic diagnosis are expensive and limited to specificity and sensitivity. Aptamers are single-stranded deoxyribonucleic acid (ssDNA) and ribonucleic acid (RNA) oligonucleotide or peptide sequence generated in vitro based on the binding affinity of aptamer-target by a process known as Systematic Evolution of Ligands by Exponential Enrichment (SELEX). By taking several advantages over monoclonal antibodies and other conventional small-molecule therapeutics, such as high specificity and affinity, negligible batch-to-batch variation, flexible modification and production, thermal stability, low immunogenicity and lack of toxicity, aptamers are presently becoming promising novel diagnostic and therapeutic agents. This review describes the prospective application of aptamerbased laboratory diagnostic assays and therapeutics for pathogenic bacteria and toxins in bloodstream infections.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Mikail Dogan ◽  
Lina Kozhaya ◽  
Lindsey Placek ◽  
Courtney Gunter ◽  
Mesut Yigit ◽  
...  

AbstractDevelopment of antibody protection during SARS-CoV-2 infection is a pressing question for public health and for vaccine development. We developed highly sensitive SARS-CoV-2-specific antibody and neutralization assays. SARS-CoV-2 Spike protein or Nucleocapsid protein specific IgG antibodies at titers more than 1:100,000 were detectable in all PCR+ subjects (n = 115) and were absent in the negative controls. Other isotype antibodies (IgA, IgG1-4) were also detected. SARS-CoV-2 neutralization was determined in COVID-19 and convalescent plasma at up to 10,000-fold dilution, using Spike protein pseudotyped lentiviruses, which were also blocked by neutralizing antibodies (NAbs). Hospitalized patients had up to 3000-fold higher antibody and neutralization titers compared to outpatients or convalescent plasma donors. Interestingly, some COVID-19 patients also possessed NAbs against SARS-CoV Spike protein pseudovirus. Together these results demonstrate the high specificity and sensitivity of our assays, which may impact understanding the quality or duration of the antibody response during COVID-19 and in determining the effectiveness of potential vaccines.


Nanomaterials ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1207
Author(s):  
Hong Jae Cheon ◽  
Quynh Huong Nguyen ◽  
Moon Il Kim

Inspired by the active site structure of natural horseradish peroxidase having iron as a pivotal element with coordinated histidine residues, we have developed histidine coated magnetic nanoparticles (His@MNPs) with relatively uniform and small sizes (less than 10 nm) through one-pot heat treatment. In comparison to pristine MNPs and other amino acid coated MNPs, His@MNPs exhibited a considerably enhanced peroxidase-imitating activity, approaching 10-fold higher in catalytic reactions. With the high activity, His@MNPs then were exploited to detect the important neurotransmitter acetylcholine. By coupling choline oxidase and acetylcholine esterase with His@MNPs as peroxidase mimics, target choline and acetylcholine were successfully detected via fluorescent mode with high specificity and sensitivity with the limits of detection down to 200 and 100 nM, respectively. The diagnostic capability of the method is demonstrated by analyzing acetylcholine in human blood serum. This study thus demonstrates the potential of utilizing His@MNPs as peroxidase-mimicking nanozymes for detecting important biological and clinical targets with high sensitivity and reliability.


2020 ◽  
Vol 47 (12) ◽  
pp. 1760-1767
Author(s):  
Sarah M. Wade ◽  
Trudy McGarry ◽  
Siobhan C. Wade ◽  
Ursula Fearon ◽  
Douglas J. Veale

ObjectiveMicroRNA (miRNA) are small endogenous regulatory RNA molecules that have emerged as potential therapeutic targets and biomarkers in autoimmunity. Here, we investigated serum miRNA levels in patients with psoriatic arthritis (PsA) and further assessed a serum miRNA signature in therapeutic responder versus nonresponder PsA patients.MethodsSerum samples were collected from healthy controls (HC; n = 20) and PsA patients (n = 31), and clinical demographics were obtained. To examine circulatory miRNA in serum from HC and PsA patients, a focused immunology miRNA panel was analyzed utilizing a miRNA Fireplex assay (FirePlex Bioworks Inc.). MiRNA expression was further assessed in responders versus nonresponders according to the European League Against Rheumatism response criteria.ResultsSix miRNA (miR-221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p, miR-26a-5p, and miR-21-5p) were significantly higher in PsA compared to HC (all P < 0.05), with high specificity and sensitivity determined by receiver-operating characteristic curve analysis. Analysis of responder versus nonresponders demonstrated higher baseline levels of miR-221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p, and miR-26a-5p were associated with therapeutic response.ConclusionThis study identified a 6-serum microRNA signature that could be attractive candidates as noninvasive markers for PsA and may help to elucidate the disease pathogenesis.


2019 ◽  
Vol 27 (3) ◽  
pp. 230949901986867 ◽  
Author(s):  
Alasdair JA Santini ◽  
Chetan A Jakaraddi ◽  
Fotis Polydoros ◽  
Sree Metikala

Postoperative urinary retention necessitating catheterization after major lower limb arthroplasty surgery adds to the patients’ postoperative discomfort and increases the risk of urinary tract infection with potential risk of transient bacteraemia and seeding of infection to prosthetic joints. Preoperative evaluation of patients with lower urinary tract symptoms may help to identify at-risk patients and the International Prostate Symptoms Score (IPSS) has been used as a screening tool to quantify the severity of symptoms in males. A prospective cohort of 303 patients undergoing total hip or knee arthroplasty was evaluated using the IPSS. Patients were categorized into three symptom groups (mild, moderate and severe based on scores of 0–7, 8–18 and greater than 18, respectively) and four age groups (<50 years, 51–60 years, 61–70 years and greater than 70 years). Twenty-six patients (8.6%) developed urinary retention and were catheterized postoperatively; of these, 16 were male and 10 were female. Statistical analysis using logistic regression models showed significant association between severe IPSS scores (>18) and urinary retention requiring catheterization in both males and females with both high specificity and sensitivity in the test in predicting postoperative catheterization. Hence, this test is a valid preoperative screen in predicting postoperative catheterization.


2013 ◽  
Vol 57 (2) ◽  
pp. 199-208 ◽  
Author(s):  
Krystyna Pohorecka ◽  
Piotr Skubida ◽  
Piotr Semkiw ◽  
Artur Miszczak ◽  
Dariusz Teper ◽  
...  

Abstract The effects to honeybee colonies (Apis mellifera L.) during and after exposure to flowering maize (Zea mays L.), grown from seeds coated with clothianidin and imidacloprid was assessed in field-realistic conditions. The experimental maize crops were adjacent to the other flowering agriculture plants. Honey bee colonies were placed in three differently protected maize fields throughout the blooming period, and thereafter they were transferred to a stationary apiary. Samples of pollen loads, bee bread, and adult bees were collected and analyzed for neonicotinoid residues. To ensure high specificity and sensitivity of detection of the analyzed pesticides, a modified QuEChERS extraction method and liquid chromatography coupled with tandem mass spectrometry were used. Clothianidin was detected only in the samples of pollen loads. Their residue levels ranged from 10.0 to 41.0 ng/g (average 27.0 ng/g). Imidacloprid was found in no investigated sample. No negative effects of neonicotinoid seed-treated maize on the development and long-term survival of honey bee colonies were observed. The low proportion of Zea mays pollen in total bee-collected pollen during the maize flowering period was noted. The findings suggest that maize plants are less attractive forage for honey bees than phacelia (Phacelia tanacetifolia Benth.), buckwheat (Fagopyrum Mill.), white clover (Trifolium repens L.), goldenrod (Solidago L.), and vegetation from Brassicaceae family. The results indicate a possibility of reducing the risk of bees being exposed to the toxic effect of insecticidal dusts dispersed during maize sowing by seeding, in the areas surrounding maize crops, plants that bloom later in the year.


2010 ◽  
Vol 63 (1-2) ◽  
pp. 47-50
Author(s):  
Sonja Vesic ◽  
Jelica Vukicevic ◽  
Eleonora Gvozdenovic ◽  
Dusan Skiljevic ◽  
Slobodanka Janosevic ◽  
...  

Introduction. Nongonococcal urethritis is the most common sexually transmitted infection in men, with vast majority of the etiological agents such as Chlamydia trachomatis, followed by urogenital mycoplasmas. The aim of this study was to determine the prevalence of Chlamydia trachomatis, Ureaplasma urealyticum and Mycoplasma hominis in nongonococcal urethritis in men, and to examine infections associated with these agents. Material and methods. 299 sexually active, heterosexual men with nongonococcal urethritis were included into the study. Urethral samples were taken with a dacron swab placed into the urethra up to 2-3 cm. The Direct immunojluorescence tehnique was performed for identification of Chlamydia trachomatis. Ureaplasma urealyticum and Mycoplasma hominis were detected with Mycoplasma 1ST assay. Results. Chlamydia trachomatis was detected in 22.75%, Uraeplasma urealyticum in 21.08% and Mycoplasma hominis in 8.02% cases. We found no significant differences in prevalence between Chlamydia trachomatis and Ureaplasma urealyticym (p>0.05). Monoinjections were found in 51.85% with significantly higher rate (p<0.01) than associated infections (11.70%). Among associated infections, coinfection of Chlamydia trahomatis and Ureaplasma urealyticum was predominant. Association of Chlamydia trachomatis with urogenital mycoplasmas was significantly higher (p<0.05) than the one between Ureaplasma urealyticum and Mycoplasma hominis. In 36.45% patients no patogenic microorganisms were detected. Conclusion. These results confirmed the etiological role of Chlamydia trachomatis and urogenital mycoplasmas in nongonococcal urethritis with prevalence of 51.85% in monoinfections and 11.70% in associated infections. In 36.45% of cases the etiology of urethritis was not elucidated. These results suggest that more sensitive diagnostic tool should be applied when searching for the detailed etiology of nongonococcal urethritis.


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