18S ribosomal DNA based PCR diagnostic assay for Trichomonas vaginalis infection in symptomatic and asymptomatic women in India

Dwivedi, Surya Prakash; Husain, Nuzhat; Rb, Singh; Malla, Nancy

doi:10.1016/s2222-1808(12)60031-0

Cited by 5 publications

(3 citation statements)

References 23 publications

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“…Plasmids were then amplified using the primers M13Forward (M13F) and M13Reverse (M13R) (Invitrogen, France) according to the manufacturer's instructions. The fragments obtained were then digested with HaeIII (Promega, USA) (Dwivedi et al, 2012). Clones were grouped according to RFLP banding patterns and further sequenced using primers M13F (5′-GTAAAACGACGGCCAG-3′) and M13R (5′-CAGGAAACAGCTATGAC-3′).…”

Section: Cloning and Sequencing Of Its Rrna Gene Amplified From The Tmentioning

confidence: 99%

Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches

Chobba¹,

Elleuch²,

Ayadi

et al. 2013

J. Zhejiang Univ. Sci. B

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Endophytic flora plays a vital role in the colonization and survival of host plants, especially in harsh environments, such as arid regions. This flora may, however, contain pathogenic species responsible for various troublesome host diseases. The present study is aimed at investigating the diversity of both cultivable and non-cultivable endophytic fungal floras in the internal tissues (roots and leaves) of Tunisian date palm trees (Phoenix dactylifera). Accordingly, 13 isolates from both root and leaf samples, exhibiting distinct colony morphology, were selected from potato dextrose agar (PDA) medium and identified by a sequence match search wherein their 18S-28S internal transcribed spacer (ITS) sequences were compared to those available in public databases. These findings revealed that the cultivable root and leaf isolates fell into two groups, namely Nectriaceae and Pleosporaceae. Additionally, total DNA from palm roots and leaves was further extracted and ITS fragments were amplified. Restriction fragment length polymorphism (RFLP) analysis of the ITS from 200 fungal clones (leaves: 100; roots: 100) using HaeIII restriction enzyme revealed 13 distinct patterns that were further sequenced and led to the identification of Alternaria, Cladosporium, Davidiella (Cladosporium teleomorph), Pythium, Curvularia, and uncharacterized fungal endophytes. Both approaches confirmed that while the roots were predominantly colonized by Fusaria (members of the Nectriaceae family), the leaves were essentially colonized by Alternaria (members of the Pleosporaceae family). Overall, the findings of the present study constitute, to the authors' knowledge, the first extensive report on the diversity of endophytic fungal flora associated with date palm trees (P. dactylifera).

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Section: Cloning and Sequencing Of Its Rrna Gene Amplified From The Tmentioning

confidence: 99%

Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches

Chobba¹,

Elleuch²,

Ayadi

et al. 2013

J. Zhejiang Univ. Sci. B

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“…The extracted DNA can be used for PCR analysis, genetic or environmental research, and human screening. The primer set Tv1 (TAA TGG CAG AAT CTT TGG AG), and Tv2 (GAA CTT TAA CCG AAG GAC TTC) was used is designed to amplify a DNA product of 312 bp, according to Dwivedi et al, [12]. After preparation of the PCR reaction (20 µl), amplification was carried out in a thermocycler (BioRad, USA Biometra).…”

Section: Design and Populationmentioning

confidence: 99%

Trichomonas vaginalis Infection and Risk Factors among Infertile Women in Gaza, Palestine

MAQADMA¹,

Al-Hindi²

2021

Journal of Microbiology and Infectious Diseases

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Objectives: Objectives: Trichomonas vaginalis is a sexually transmitted disease and can infect a wide range of human beings. The present study was conducted to determine the prevalence and risk factors associated with Trichomoniasis infection among infertile women in Gaza city, Palestine.Methods: A descriptive-analytical cross-sectional study was conducted between December 2013 to April 2014 in Al Basma medical center in Gaza city. Ethical approval was obtained from the Helsinki Committee in Gaza to conduct the study, and all participants gave consent. A total of 120 endocervical swabs were collected from females attending the center to manage delayed conception. The samples were processed using the PCR technique with Tv1-Tv2 primers in Islamic Universities laboratories. In addition, a structured questionnaire was conducted with all participants regarding socio-demographic data, risk factors, and symptomatology. Results:The prevalence of T. vaginalis was 5.8% among the participants. There was a significant relation between infection and age of the patients, marriage age, the number of previous vaginal infections, and inguinal erythema (p=0.01). A higher infection rate was found among unemployed, low educated, mid-zone residents, and live in crowded houses, and married women with smoker husbands. Conclusion:We concluded that the prevalence of Trichomoniasis is not attributed mainly to personal hygiene, so in future studies, the examination tools used in the clinics should be included /tested to sort the source of infection.

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“…However, current detection methods do not meet these requirements, as the infection is difficult to diagnose clinically, since the associated symptoms may also be present in other urogenital infections and sexually transmitted diseases. , Current diagnostic methods available for T. vaginalis detection include direct visualization of the trophozoite by wet mount microscopy, culture methods, immunoassay techniques, and nucleic acid amplification tests (NAAT). On the one hand, the most widely used method, the wet mount microscopy, is rapid and inexpensive but suffers from a low sensitivity of only 38–82%. ,, The culture method, on the other hand, is considered as the gold standard but requires at least 2–7 d of incubation. , While NAAT methods, such as polymerase chain reaction (PCR) amplification, provide highly reliable results, − they require expensive equipment, reagents, infrastructure, and trained personnel.…”

mentioning

confidence: 99%

Aptamer Selection against a Trichomonas vaginalis Adhesion Protein for Diagnostic Applications

Espiritu¹,

Justo²,

Jauset‐Rubio

et al. 2018

ACS Infect. Dis.

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Trichomoniasis, caused by Trichomonas vaginalis, is the leading nonviral sexually transmitted infection worldwide. We report the selection of a DNA aptamer against a T. vaginalis adhesion protein, AP65, using a microtiter plate-based in vitro combinatorial chemistry process termed systematic evolution of ligands by exponential enrichment. The enriched library pool was sequenced by next-generation sequencing, and several aptamer candidates with high affinity and specificity were identified. The aptamer with the highest affinity and specificity had a K in the low nanomolar range, as confirmed by three different techniques: surface plasmon resonance, enzyme-linked aptamer assay, and biolayer interferometry. The selected aptamer was demonstrated to have a high specificity to the AP65 protein and to T. vaginalis cells with no cross-reactivity to other enteric and urogenital microorganisms. Current work is focused on the development of inexpensive and easy-to-use aptamer-based diagnostic assays for the reliable and rapid detection of T. vaginalis in vaginal swabs.

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18S ribosomal DNA based PCR diagnostic assay for Trichomonas vaginalis infection in symptomatic and asymptomatic women in India

Cited by 5 publications

References 23 publications

Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches

Fungal diversity in adult date palm (Phoenix dactylifera L.) revealed by culture-dependent and culture-independent approaches

Trichomonas vaginalis Infection and Risk Factors among Infertile Women in Gaza, Palestine

Aptamer Selection against a Trichomonas vaginalis Adhesion Protein for Diagnostic Applications

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