scholarly journals Identification of Mycoplasma genitalium from clinical swabs by direct PCR

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1993
Author(s):  
Robinson M. Irekwa ◽  
Perpetual Ndung'u ◽  
Peter Kipkemboi ◽  
Tonny Teya ◽  
Anne Wanjiru Mwangi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Workers ◽  
Genital Tract ◽  
Mycoplasma Genitalium ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Direct Pcr ◽  
The World ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

Mycoplasma genitalium is one of the smallest self-replicating organisms. It is an obligate parasite found in the human genital tract. In men, the bacteria cause both acute and chronic non-gonococcal urethritis (NGU). In women, it has been associated with pelvic inflammatory disease and cervicitis among other related infections. Treatment of M. genitalium related infections has been effective using antibiotics such as the macrolides (e.g. azithromycin) and fluoroquinolones. However, there have been recorded cases of resistance to these antibiotics in various parts of the world as a result of a mutation in the 23SrRNA gene, although the antibiotic resistance has not been well established. The aim of this study was to detect M. genitalium in 352 swab samples collected from a clinic for sex workers in Nairobi, Kenya. DNA was extracted from the swabs and stored as a crude extract at -31°C. The swab lysates were subjected to direct polymerase chain reaction using primers that specifically target the 16S rRNA gene for M. genitalium. A total of 29 samples tested positive for M. genitalium. The data results showed a M. genitalium prevalence of 8.24% among sex workers in Nairobi, Kenya.

Sexually transmissible infections among female sex workers in Manado, Indonesia, using a multiplex polymerase chain reaction-based reverse line blot assay

Sexual Health ◽  
2011 ◽  
Vol 8 (1) ◽  
pp. 52 ◽  
Author(s):  
Ferra O. Mawu ◽  
Stephen C. Davies ◽  
Michelle McKechnie ◽  
Endang R. Sedyaningsih ◽  
Asti Widihastuti ◽  
...  
Keyword(s):  
Risk Factors ◽  
Polymerase Chain Reaction ◽  
Sex Work ◽  
Sex Workers ◽  
Multiplex Polymerase Chain Reaction ◽  
Female Sex Workers ◽  
Mycoplasma Genitalium ◽  
Chain Reaction ◽  
Sexually Transmissible Infections ◽  
Polymerase Chain

Background: Sexually transmissible infections (STIs) remain highly prevalent, and HIV is increasing, among female sex workers (FSWs) in Indonesia. Our aim was to determine the prevalence of, and risk factors for, STIs among FSWs in Manado, Indonesia. Methods: We recruited FSWs mainly at their workplace: they completed a questionnaire and provided a urine sample and self-collected vaginal swab. Samples were tested using multiplex polymerase chain reaction, followed by reverse line blot hybridisation. Results: We recruited 221 FSWs, (median age: 25 years). During the previous 3 months, 30% reported never using condoms; only 2.7% always used condoms. Of 217 women with urine samples, 49% had a ‘curable STI’: 10.6% with gonorrhoea, 26.7% with chlamydia, 12.4% with Mycoplasma genitalium and 22.6% with trichomoniasis. Independent risk factors for gonorrhoea were: domiciled outside North Sulawesi (P = 0.001) and age 16–25 years (P = 0.02); for chlamydia: no prior history of STI symptoms (P = 0.003) and age 16–25 years (P = 0.02); for Mycoplasma genitalium: number of clients on last day of sex work (P = 0.004); for trichomoniasis: number of clients per week (P = 0.04). When these four infections were grouped as any ‘curable STI’, independent associations were: number of clients on the last day of sex work (P = 0.001), age 16–25 years (P = 0.02) and sex working for fewer than 2 years (P = 0.03). Conclusions: This is the first report of M. genitalium infection in Indonesia. The high prevalence of STIs and low condom use among these FSWs suggest their vulnerability to the HIV epidemic in Indonesia. They need enhanced interventions, including outreach screening, and periodic presumptive treatment.

scholarly journals Detection of Acidovorax avenae subsp. citrulli in Watermelon Seed Using Immunomagnetic Separation and the Polymerase Chain Reaction

Plant Disease ◽  
2000 ◽  
Vol 84 (4) ◽  
pp. 470-474 ◽  
Author(s):  
R. R. Walcott ◽  
R. D. Gitaitis
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Amplification ◽  
Fold Increase ◽  
Immunomagnetic Separation ◽  
Detection Sensitivity ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Direct Pcr ◽  
Watermelon Seed ◽  
Polymerase Chain

An immunomagnetic separation and polymerase chain reaction (IMS-PCR)-based assay was developed for detecting Acidovorax avenae subsp. citrulli in watermelon seed. IMS yielded a 10-fold increase in recovery of A. avenae subsp. citrulli over direct spread-plating on King's Medium B; however, the presence of seed debris reduced IMS efficiency. Synthetic oligonucleotide primers were designed based on the 16S rRNA gene of a known A. avenae subsp. citrulli strain and tested for specific DNA amplification by PCR. The primers amplified DNA from all A. avenae subsp. citrulli strains tested but also yielded amplicons with several closely related bacteria. IMS-PCR resulted in a 100-fold increase in A. avenae subsp. citrulli detection sensitivity over direct PCR and was unaffected by PCR inhibitors in watermelon seed. The threshold of A. avenae subsp. citrulli detection for IMS-PCR was 10 CFU/ml in watermelon seed wash, and seedlots with 0.1% infestation were consistently detected. IMS-PCR represents an efficient and sensitive approach to detecting A. avenae subsp. citrulli in watermelon seedlots.

scholarly journals Molecular detection of Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania

2020 ◽  
Author(s):  
Noel Gahamanyi ◽  
Leonard E.G. Mboera ◽  
Mecky I. Matee ◽  
Dieudonné Mutangana ◽  
Raghavendra G. Amachawadi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Faecal Samples ◽  
Rrna Sequencing ◽  
Polymerase Chain ◽  
Campylobacter Species ◽  
The 16S Rrna Gene

Abstract Background: A growing number of Campylobacter species other than C. jejuni and C. coli have been considered as emerging human and animal pathogens. However, the contribution of these species to human gastroenteritis is poorly documented. This study aimed at detecting Campylobacter species from human and cattle faecal samples in Kilosa district, Tanzania using Polymerase Chain Reaction (PCR) amplification of the 16S rRNA gene, and Sanger sequencing . Methods: A total number of 100 faecal samples (70 from human and 30 from cattle) were collected from diarrheic and non-diarrheic patients and healthy cattle in Kilosa district, Tanzania from July to October 2019. Species identification was conducted by PCR and 16S rRNA sequencing. The phylogenetic analysis was carried out by comparison of the 16S rRNA gene sequences to reference strains by the Neighbor-Joining method in MEGA X. Results: Campylobacter species detection rate by PCR was 65.7% (46/70) and 20% (6/30) in humans and cattle, respectively. There were five human diarrheic cases, four showed Campylobacter presence and two were from children ≤15 years of age. In humans, the 16S rRNA sequencing revealed that C. concisus was the most predominant species occurring at a frequency of 37.8% (14/37), followed by uncultured Campylobacter spp. 24.3% (9/37) and C. hominis 21.6% (8/37). The least represented species were C. jejuni and C. lanienae all occurring at 2.7% (1/37). In cattle, five (100%) sequenced PCR products matched with C. lanienae . Phylogenetic analysis revealed that Campylobacter 16S rRNA sequences were closely related to C. concisus , uncultured Campylobacter sp., C. hominis , and C. gracilis . Conclusion: The non- C. jejuni / C. coli species are present in human and cattle faecal samples and their true occurrence is probably under-reported due to shortcomings of conventional techniques used in most diagnostic microbiology laboratories. Based on our findings, we recommend that molecular techniques be adopted for direct detection of Campylobacter species during routine laboratory screening and surveillance studies. Keywords: Campylobacter , molecular diagnostics, polymerase chain reaction, sequencing, gastroenteritis, Tanzania

scholarly journals Detection of the Legionnaires’ Disease Agent in Patients With Respiratory Symptoms by Culture, Detection of Urinary Antigen and Polymerase Chain Reaction of the 16S rRNA Gene in Ahvaz, Iran

2017 ◽  
Vol In press (In press) ◽  
Author(s):  
Mojtaba Moosavian ◽  
Mahtab Khoshkholgh Sima ◽  
Maryam Haddadzadeh Shoushtari ◽  
Seyed Mohammad Alavi ◽  
Mohammad Amin Fazeli Naserabad ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
Respiratory Symptoms ◽  
Rrna Gene ◽  
Urinary Antigen ◽  
Chain Reaction ◽  
Disease Agent ◽  
Legionnaires Disease ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

scholarly journals Detection and genetic characterization of "Candidatus Mycoplasma haemomacaque" infection among long-tailed macaques (Macaca fascicularis) in Thailand using broad-range nested polymerase chain reaction assay

2021 ◽  
Vol 14 (4) ◽  
pp. 943-948
Author(s):  
Wanat Sricharern ◽  
Supakarn Kaewchot ◽  
Sarawan Kaewmongkol ◽  
Natnaree Inthong ◽  
Thitichai Jarudecha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Genetic Characterization ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Blood Samples ◽  
Polymerase Chain ◽  
The 16S Rrna Gene

Background and Aim: Hemoplasmas are defined as small, epicellular parasitic bacteria that can infect the red blood cells of several mammalian species. Diseases caused by these bacteria range from asymptomatic infections to acute hemolytic anemia. However, data on hemoplasmas in non-human primates in Thailand remain to be limited. Therefore, this study aims to determine the occurrence and genetic diversity of hemoplasmas among long-tailed macaques in Thailand. Materials and Methods: Blood samples were collected from 339 long-tailed macaques in three provinces of Thailand. DNA was then extracted from the blood samples and tested for hemoplasma using broad-range nested polymerase chain reaction (PCR) based on the 16S rRNA gene. PCR-positive samples were sequenced, and phylogenetic analysis for species identification was conducted. Results: In total, 38 (11.2%) out of the 339 samples were found to be positive for hemoplasmas, based on the broad-range nested PCR assay of the 16S rRNA gene. The 16S rRNA sequences of Mycoplasma spp. were highly similar (98-99% identity) to "Candidatus Mycoplasma haemomacaque." Furthermore, phylogenetic analysis using maximum likelihood demonstrated that the sequences were located in the same cluster of "Ca. M. haemomacaque." Conclusion: The detection of hemoplasmas among long-tailed macaques in Thailand is reported. Genetic characterization confirmed that these hemoplasmas are closely related to "Ca. M. haemomacaque." These results indicate that long-tailed macaques in several locations in Thailand may be infected and serve as reservoirs for this parasite.

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