scholarly journals 18S Ribosomal DNA-Based PCR for Diagnosis ofTrichomonas vaginalis

2000 ◽  
Vol 38 (7) ◽  
pp. 2683-2687 ◽  
Author(s):  
Holger Mayta ◽  
Robert H. Gilman ◽  
Maritza M. Calderon ◽  
Aren Gottlieb ◽  
Giselle Soto ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Trichomonas Vaginalis ◽  
High Sensitivity ◽  
18S Rrna Gene ◽  
Vaginal Swab ◽  
Urine Samples ◽  
Rrna Gene ◽  
Sexually Transmitted ◽  
Immunodeficiency Virus ◽  
Pcr Products

Trichomonas vaginalis remains the most common sexually transmitted parasite in the world and is considered a major risk factor in the transmission of the human immunodeficiency virus. A PCR technique using primers targeting a specific region of the 18S rRNA gene of T. vaginalis was developed. The PCR test was standardized using 15 reference strains, giving a single product of 312 bp in all strains. No amplification was observed when DNA from related organisms or human DNA was used as a target. The test was evaluated on 372 vaginal swab specimens and 361 urine samples from women attending infertility and obstetric clinics at two separate hospitals in Lima, Peru. Compared to T. vaginalis culture, the overall sensitivity and specificity of PCR of vaginal swab samples was 100% and 98%, respectively. The PCR of urine samples was 100% sensitive and 99.7% specific compared to culture of vaginal swab, but the sensitivity drops to 83.3% when compared to PCR of vaginal swabs. All culture-positive samples were found to be positive by PCR in either urine or vaginal secretion. None of the PCR-negative samples were positive by culture. The origin of the amplification was confirmed by digestion of PCR products with HaeIII. This PCR assay, which is easy to perform and has a high sensitivity and specificity, should be useful for routine diagnosis of T. vaginalisinfection.

scholarly journals Detection of Trichomonosis in Vaginal and Urine Specimens from Women by Culture and PCR

2000 ◽  
Vol 38 (10) ◽  
pp. 3585-3588 ◽  
Author(s):  
Lisa F. Lawing ◽  
Spencer R. Hedges ◽  
Jane R. Schwebke
Keyword(s):  
Sexually Transmitted Diseases ◽  
Sensitivity And Specificity ◽  
Trichomonas Vaginalis ◽  
Dna Amplification ◽  
Sexually Transmitted ◽  
Specific Pcr ◽  
Virus Acquisition ◽  
Immunodeficiency Virus ◽  
Culture Positive ◽  
Urine Specimens

Vaginal trichomonosis is a highly prevalent infection which has been associated with human immunodeficiency virus acquisition and preterm birth. Culture is the current “gold standard” for diagnosis. As urine-based testing using DNA amplification techniques becomes more widely used for other sexually transmitted diseases (STDs) such as gonorrhea and chlamydia, a similar technique for trichomonosis would be highly desirable. Women attending an STD clinic for a new complaint were screened for Trichomonas vaginalis by wet-preparation (wet-prep) microscopy and culture and for the presence of T. vaginalis DNA by specific PCR of vaginal and urine specimens. The presence of trichomonosis was defined as the detection of T. vaginalis by direct microscopy and/or culture from either vaginal samples or urine. The overall prevalence of trichomonosis in the population was 28% (53 of 190). The sensitivity and specificity of PCR using vaginal samples were 89 and 97%, respectively. Seventy-four percent (38 of 51) of women who had a vaginal wet prep or vaginal culture positive for trichomonads had microscopic and/or culture evidence of the organisms in the urine. Two women were positive for trichomonads by wet prep or culture only in the urine. The sensitivity and specificity of PCR using urine specimens were 64 and 100%, respectively. These results indicate that the exclusive use of urine-based detection of T. vaginalis is not appropriate in women. PCR-based detection of T. vaginalis using vaginal specimens may provide an alternative to culture.

EVALUATING THE RELIABILITY OF ELISA TO DETECT THE ANTIBODY AGAINST T. VAGINALIS, AND THE PREVALENCE OF T.VAGINALIS INFECTION IN HUE CITY

2013 ◽  
pp. 25-34
Author(s):  
Nu Phuong Anh Ton ◽  
Minh Chau Ngo ◽  
Phuoc Vinh Nguyen ◽  
Luigi Fiori Pier ◽  
Minh Tam Le ◽  
...  
Keyword(s):  
Antibody Response ◽  
Sensitivity And Specificity ◽  
Trichomonas Vaginalis ◽  
Transmitted Disease ◽  
High Sensitivity ◽  
Sex Behavior ◽  
Unsafe Sex ◽  
Healthy Men ◽  
Sexually Transmitted ◽  
Asymptomatic Women

Objective:The protist Trichomonas vaginalis is the most common non-viral, curable, sexually transmitted disease agent worldwide. The objective of this study is to determine the prevalence of trichomoniasis patients in Hue City, Vietnam and its serological patterns. Materials and methods: The study included 249 symptomatic women, 534 asymptomatic women, 38 healthy men, and 50 sera of children 2-10 years of age from Hue City, Vietnam from September 2010 to June 2012. In addition, specific anti - T. vaginalis antibody response was studied in a group of 46 women affected by trichomoniasis and 8 male sexual partners. All women were subjected to standard clinical examination and vaginal samples were collected for identification of Trichomonas vaginalis by wet mount and cultivation in specific media. Sera from trichomoniasis patients were used to set up immunoenzymatic techniques to detect specific antibody response for seroepidemiological studies. Results: The sensitivity and specificity of ELISA assay were 93.48%, 84.88% respectively. The prevalence of trichomoniasis diagnosed by microscopic examination in symptomatic women and asymptomatic groups were 19.3% (42/243, 95% CI = 12.8% - 22.7%) and 0.7% (4/534, 95% CI = 0.18% - 1.8%), respectively. The seroprevalence from general population were found 18.9% in women and 8.7% in men. The seroprevalence were 31.3% in symptomatic women, 13.3% in asymptomatic women. The seroprevalence was 14% in safe sex behavior women to compare with 22.7% in unsafe sex behavior women. There were 7.9% seropositive from sera of healthy men and 12.5% seropositive from sera of men partners of trichomoniasis women. Conclusion: In general, the prevalence of T. vaginalis infection is high in symptomatic women and low in asymptomatic women. ELISA essay yielded high sensitivity and specificity in diagnosis of vaginal trichomoniasis. Key words: T. vaginalis, seroepidemiology, ELISA.

scholarly journals Improved molecular laboratory productivity by consolidation of testing on the new random-access analyzer Alinity m

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Martin Obermeier ◽  
Monia Pacenti ◽  
Robert Ehret ◽  
Francesco Onelia ◽  
Rory Gunson ◽  
...  
Keyword(s):  
Trichomonas Vaginalis ◽  
Random Access ◽  
Mycoplasma Genitalium ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
B Virus ◽  
Immunodeficiency Virus ◽  
International Multicenter Study ◽  
Laboratory Productivity ◽  
Laboratory Workflow

AbstractObjectivesAutomated molecular analyzers have accelerated diagnosis, allowing earlier intervention and better patient follow-up. A recently developed completely automated molecular analyzer, Alinity™ m (Abbott), offers consolidated, continuous, and random-access testing that may improve molecular laboratory workflow.MethodsAn international, multicenter study compared laboratory workflow metrics across various routine analyzers and Alinity m utilizing assays for human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), hepatitis B virus (HBV), high-risk human papillomavirus (HR HPV), and sexually transmitted infection (STI) (Chlamydia trachomatis [CT]/Neisseria gonorrhoeae [NG]/Trichomonas vaginalis [TV]/Mycoplasma genitalium [MG]). Three turnaround times (TATs) were assessed: total TAT (sample arrival to result), sample onboard TAT (sample loading and test starting to result), and processing TAT (sample aspiration to result).ResultsTotal TAT was reduced from days with routine analyzers to hours with Alinity m, independent of requested assays. Sample onboard TATs for standard workflow using routine analyzers ranged from 7 to 32.5 h compared to 2.75–6 h for Alinity m. The mean sample onboard TAT for STAT samples on Alinity m was 2.36 h (±0.19 h). Processing TATs for Alinity m were independent of the combination of assays, with 100% of results reported within 117 min.ConclusionsThe consolidated, continuous, random-access workflow of Alinity m reduces TATs across various assays and is expected to improve both laboratory operational efficiency and patient care.

A Review on Diagnostic Methods for Trichomonas Vaginalis

2021 ◽  
Author(s):  
Fatemeh Rahmani ◽  
Yahya Ehteshaminia ◽  
Hamid Mohammadi ◽  
Seif Ali Mahdavi
Keyword(s):  
Trichomonas Vaginalis ◽  
Pap Smear ◽  
High Sensitivity ◽  
Protozoan Parasite ◽  
Signs And Symptoms ◽  
Diagnostic Methods ◽  
Culture Methods ◽  
Transmitted Infection ◽  
Sexually Transmitted ◽  
Asymptomatic Individuals

Introduction: Trichomoniasis is the most common non-viral sexually transmitted infection in the world, caused by the protozoan parasite Trichomonas vaginalis, which infects the urogenital tract of men and women. Approximately, 250 million new cases of Trichomonas vaginalis Infection are reported worldwide each year. Trichomoniasis is also considered an important HIV co-infection. The infection is often asymptomatic but can be accompanied by symptoms such as severe inflammation, itching and irritation, foamy discharge, and malodorous smell mucus, but the signs and symptoms of the disease are not sufficient for specific diagnosis. Material and Methods: In this study, the websites of PubMed, Google Scholar, SID, and Margiran were searched and related articles were reviewed. Results: Only screening and the use of highly sensitive and specific diagnostic methods can identify asymptomatic individuals. Today, the most common way to diagnose the infection is to use wet slide, Pap smear and culture methods that do not have high sensitivity and specificity. Also, due to the increase in infection and its complications, finding an efficient, rapid, and easy test to detect the parasite and differentiate Trichomoniasis vaginitis from other sexually transmitted diseases is considered important and necessary. Conclusion: Nowadays, there are several diagnostic methods that differentiate trichomoniasis infection from other sexually transmitted infections with high accuracy and sensitivity. Of course, existing diagnostic methods mostly use women's urine and vaginal samples for diagnosis, and methods that specifically diagnose the infection in men are more limited.

scholarly journals Emergence of Babesia Conradae Infection in Coyote-hunting Greyhounds in Oklahoma, USA

2021 ◽  
Author(s):  
Erin M Stayton ◽  
Megan Lineberry ◽  
Jennifer Thomas ◽  
Tina Bass ◽  
Kelly Allen ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
18S Rrna Gene ◽  
Post Treatment ◽  
Rrna Gene ◽  
Babesia Gibsoni ◽  
Wide Range ◽  
Pcr Products ◽  
Life Threatening ◽  
Tick Vectors

Abstract Background: Babesia species are intraerythrocytic Apicomplexan parasites that infect a wide range of vertebrate hosts. These pathogens are typically transmitted either by tick vectors or by direct blood-to-blood contact, and may cause life-threatening clinical disease such as thrombocytopenia, hemolytic anemia, and acute renal failure in canine hosts. While Babesia vogeli and Babesia gibsoni infections have both been reported in Oklahoma, reports of B. conradae infections have been limited to California. Methods: Whole blood samples were collected in EDTA tubes from all dogs in four separate kennels in Oklahoma. DNA was extracted from each blood sample and a nested PCR was performed using general Apicomplexan primers for the partial 18S rRNA gene. PCR products were electrophoresed in agarose matrix and appropriately sized amplicons were sequenced. Sequences were compared to reference 18S rRNA sequences available in GenBank, and samples with >98% homology to B. conradae (GenBank MK256976) were considered positive. B. conradae positive dogs were then treated with atovaquone (13.5 mg/kg TID) and azithromycin (10 mg/kg SID) for 10 days and retested at 30 and 60 days post treatment by PCR. Results: Fifteen of 40 dogs tested positive for B. conradae with 98–100% sequence homology to B. conradae from California. All positive cases were coyote-hunting Greyhounds. Treatment of clinically ill dogs with atovaquone and azithromycin resulted in complete clinical recovery in clinically ill dogs and all treated dogs had negative follow-up PCR at 30 and 60 days post treatment. Conclusions: Collectively, this study (i) documents the occurrence of B. conradae in Oklahoma, (ii) highlights this pathogen as a differential to be considered when clinical signs are present, and (iii) supports the use of atovaquone and azithromycin as effective treatment in these cases.

scholarly journals Stenamoeba dejonckheerei sp. nov., a Free-Living Amoeba Isolated from a Thermal Spring

Pathogens ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 586
Author(s):  
Manuel Alejandro Borquez-Román ◽  
Luis Fernando Lares-Jiménez ◽  
Libia Zulema Rodriguez-Anaya ◽  
Jose Reyes Gonzalez-Galaviz ◽  
Paul A. Fuerst ◽  
...  
Keyword(s):  
Thermal Spring ◽  
Small Subunit ◽  
18S Rrna Gene ◽  
Rrna Gene ◽  
Likelihood Analysis ◽  
Ribosomal Ribonucleic Acid ◽  
Pcr Products ◽  
Northwestern Mexico ◽  
Rrna Gene Sequencing ◽  
A New Species

Two amoeboid organisms were obtained from water samples taken from a thermal spring, "Agua Caliente", in Northwestern Mexico. The isolates were obtained when samples were cultivated at 37 °C on non-nutrient agar coated with Escherichia coli. The initial identification of the isolates was performed morphologically using light microscopy. The samples were found to have trophozoite morphology consistent with members of the genus Stenamoeba, a genus derived in 2007 from within the abolished polyphyletic genus Platyamoeba. Further analysis was performed by sequencing PCR products obtained using universal eukaryotic primers for the small subunit ribosomal ribonucleic acid (SSU rRNA) gene. Sequencing primers were designed to allow the comparison of the 18S rRNA gene sequences of the new isolates with previous sequences reported for Stenamoeba. Phylogenetic relationships among sequences from Stenamoeba were determined using Maximum Likelihood analysis. The results showed the two "Agua Caliente" sequences to be closely related, while clearly separating them from those of other Stenamoeba taxa. The degrees of sequence differentiation from other taxa were considered sufficient to allow us to propose that the Mexican isolates represent a new species.

scholarly journals Evaluation of the Determine Rapid Syphilis TP Assay Using Sera

2004 ◽  
Vol 11 (1) ◽  
pp. 98-101 ◽  
Author(s):  
Theresa Diaz ◽  
Maria de Gloria Bonecini Almeida ◽  
Ingebourg Georg ◽  
Suely de Carvalho Maia ◽  
Rogerio Valls de Souza ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Treponema Pallidum ◽  
High Sensitivity ◽  
Hiv Positive ◽  
Hemagglutination Assay ◽  
Laboratory Facilities ◽  
Immunodeficiency Virus ◽  
Resource Poor ◽  
Clinical Conditions ◽  
Hiv Negative

ABSTRACT The Abbott Determine Rapid Syphilis TP assay is a treponemal test that can be used in resource-poor settings that lack laboratory facilities. However, this test has not been extensively evaluated. We measured its sensitivity and specificity by using stored serum specimens (n = 567) from all persons who tested Treponema pallidum hemagglutination assay (TPHA) positive (n = 250) or TPHA indeterminate (n = 17) in the year 2001 and the first 300 patients in 2001 who tested TPHA negative at the Evandro Chagas Research Institute in Rio de Janeiro, Brazil. This rapid assay was independently interpreted by three different observers. With TPHA results as the reference, sensitivity ranged between readers from 95.6 to 98.4% and specificity ranged from 97.3 to 95.7%. There was little interreader variability in the interpretation of results, with approximately 98% agreement for all reader combinations. Of samples from persons with human immunodeficiency virus (HIV) infection (n = 198), sensitivity was 96.9 to 99.2% and it was 94.4 to 96.3% among HIV-negative persons (n = 127). Specificity was 92.4 to 95.5% among HIV-positive persons and 97.2 to 100% among HIV-negative persons. We found this test to have high sensitivity and specificity and little interreader variability, indicating that it may be easily used in resource-poor settings without laboratory facilities. Further studies are needed using this test on whole blood and under the clinical conditions for which it is intended.

scholarly journals DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen

BioTechniques ◽  
2020 ◽  
Vol 69 (6) ◽  
pp. 450-454
Author(s):  
Lara Dutra ◽  
Ole Franz ◽  
Veli-Mikko Puupponen ◽  
Marja Tiirola
Keyword(s):  
Phase Separation ◽  
Liquid Nitrogen ◽  
Sensitivity And Specificity ◽  
Dna Content ◽  
High Sensitivity ◽  
Droplet Microfluidics ◽  
Rapid Freezing ◽  
Dna Recovery ◽  
Pcr Products ◽  
Pcr Assays

Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a simple and environmentally friendly protocol for recovering DNA from ddPCR.

scholarly journals In vitro effect of tinidazole and furazolidone on metronidazole-resistant Trichomonas vaginalis.

1996 ◽  
Vol 40 (5) ◽  
pp. 1121-1125 ◽  
Author(s):  
E M Narcisi ◽  
W E Secor
Keyword(s):  
Trichomonas Vaginalis ◽  
Drug Efficacy ◽  
Protozoan Parasite ◽  
Enteric Bacteria ◽  
Sexually Transmitted ◽  
Drug Of Choice ◽  
Immunodeficiency Virus ◽  
Placental Membranes ◽  
Vitro Effect

Trichomonas vaginalis is a common sexually transmitted protozoan parasite. Although often considered simply a nuisance infection, T. vaginalis has been implicated in premature rupture of placental membranes and increases in the risk of acquiring human immunodeficiency virus. Metronidazole, a 5-nitroimidazole, is currently the drug of choice to treat T. vaginalis infection. Because some patients have severe reactions to metronidazole and others are infected with metronidazole-resistant T. vaginalis, we were prompted to investigate alternative therapies. Tinidazole, another 5-nitroimidazole used in other countries to treat T. vaginalis infections, and furazolidone, a nitrofuran presently used to treat giardiasis and infections with some anaerobic enteric bacteria, were investigated for effectiveness against 9 metronidazole-susceptible and 12 metronidazole-resistant T. vaginalis patient isolates. The in vitro aerobic and anaerobic minimum lethal concentrations (MLC) and the time for drug efficacy were determined. Tinidazole killed the metronidazole-susceptible isolates at a low MLC but was effective against only 4 of the 12 metronidazole-resistant isolates. In contrast, furazolidone was effective at a low MLC for all isolates. When tinidazole was effective, it required > 6 h to kill trichomonads. However, furazolidone killed both metronidazole-susceptible and resistant trichomonads within 2 to 3 h of exposure. These data suggest that furazolidone may be a good candidate for treating metronidazole-resistant trichomoniasis and that further investigation of this drug is warranted.

scholarly journals Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis screening and treatment of pregnant women in Port-au-Prince, Haiti

2017 ◽  
Vol 28 (11) ◽  
pp. 1130-1134 ◽  
Author(s):  
Claire C Bristow ◽  
Patricia Mathelier ◽  
Oksana Ocheretina ◽  
Daphne Benoit ◽  
Jean W Pape ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Neisseria Gonorrhoeae ◽  
Pregnant Women ◽  
Laboratory Testing ◽  
Trichomonas Vaginalis ◽  
Opportunistic Infections ◽  
Newborn Health ◽  
Vaginal Swab ◽  
Sexually Transmitted ◽  
High Prevalence

In Haiti, routine screening for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Trichomonas vaginalis (TV) among pregnant women is not conducted; yet these sexually transmitted infections (STIs) are associated with adverse birth and newborn health outcomes. We aimed to assess the acceptability and feasibility of screening and the prevalence of STIs among pregnant women in Port-au-Prince, Haiti. Pregnant women of at least 18 years of age who attend Haitian Study Group for Kaposi’s sarcoma and Opportunistic Infections (GHESKIO) clinics in Port-au-Prince, Haiti provided self-collected vaginal swab specimens. Laboratory testing was done with Xpert® CT/NG and Xpert® TV. The results of this study showed that of the 322 pregnant women who visited GHESKIO for their regular scheduled appointments, 300 (93.2%) consented for CT, NG, and TV testing. Of those, 107 women (35.7%) tested positive for at least one STI. There were 42 (14.7%) cases of CT, 8 (2.8%) NG, and 83 (29.0%) TV infections. Most infections were treated – 122 of 133 (91.7%). In summary, we found that it was highly acceptable and feasible to implement CT, NG, and TV screening among pregnant women in Port-au-Prince, Haiti. We found high prevalence of STIs among pregnant women, which suggest that STI screening in this population may be warranted.

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