Prevalence and Genotyping of Trichomonas vaginalis Infected to dsRNA Virus by PCR–Restriction Fragment Length Poly-morphism (RFLP)

Orujzadeh, Fariba; Tabatabaie, Fatemeh; Khanaliha, Khadijeh; Akhlaghi, Lame; Bokharaei‐Salim, Farah; Fallah, Soudabeh; Esteghamati, Abdoulreza; Masoumi-Asl, Hossein

doi:10.18502/ijpa.v14i2.1137

Cited by 4 publications

(5 citation statements)

References 29 publications

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“…We discovered that the clinical samples of T. vaginalis were genotyped E, M or N. In some previous studies, genotypes E and G were reported as the most prevalent, but they were detected in both TVV and TVV −/− samples, while the distribution of virus-carrying strains in the phylogenetic tree was random and not in the same cluster. This suggests that TVV may infect virus-free strains and that there is no relationship between the presence or absence of virus in the parasite and the genealogical status of the parasite [ 31 ]. In addition, we found a strain of T. vaginalis carrying the virus, which implied that the treatment and control of this T. vaginalis strain may be more difficult than those of the T. vaginalis -only strain.…”

Section: Discussionmentioning

confidence: 99%

Establishment and application of a CRISPR-Cas12a-based RPA-LFS and fluorescence for the detection of Trichomonas vaginalis

Wang

et al. 2022

Parasites Vectors

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Background Infection with Trichomonas vaginalis can lead to cervicitis, urethritis, pelvic inflammatory disease, prostatitis and perinatal complications and increased risk of HIV transmission. Here, we used an RPA-based CRISPR-Cas12a assay system in combination with a lateral flow strip (LFS) (referred to as RPA-CRISPR-Cas12a) to establish a highly sensitive and field-ready assay and evaluated its ability to detect clinical samples. Methods We developed a one-pot CRISPR-Cas12a combined with RPA-based field detection technology for T. vaginalis, chose actin as the target gene to design crRNA and designed RPA primers based on the crRNA binding site. The specificity of the method was demonstrated by detecting genomes from nine pathogens. To improve the usability and visualize the RPA-CRISPR-Cas12a assay results, both fluorescence detection and LFS readouts were devised. Results The RPA-CRISPR-Cas12a assay platform was completed within 60 min and had a maximum detection limit of 1 copy/µl and no cross-reactivity with Candida albicans, Mycoplasma hominis, Neisseria gonorrhoeae, Escherichia coli, Cryptosporidium parvum, G. duodenalis or Toxoplasma gondii after specificity validation. Thirty human vaginal secretions were tested by RPA-CRISPR-Cas12a assays, and the results were read by a fluorescent reporter and LFS biosensors and then compared to the results from nested PCR detection of these samples. Both RPA-CRISPR-Cas12a assays showed 26.7% (8/30) T. vaginalis-positive samples and a consistency of 100% (8/8). The RPA-CRISPR-Cas12a assays had a higher sensitivity than nested PCR (only seven T. vaginalis-positive samples were detected). Conclusions The T. vaginalis RPA-CRISPR-Cas12a assay platform in this study can be used for large-scale field testing and on-site tests without the need for trained technicians or costly ancillary equipment. Graphical abstract

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Section: Discussionmentioning

confidence: 99%

Establishment and application of a CRISPR-Cas12a-based RPA-LFS and fluorescence for the detection of Trichomonas vaginalis

Wang

et al. 2022

Parasites Vectors

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“…At first, wet mount samples with sterile phosphate-buffered saline(PBS; pH 7.3 ± 0.1) were prepared and observed using light microscope with 10X and then 40X magnification and then another swab was inoculated into a culture tube at 37°C in Diamond’s TYM medium with 10% heat-inactivated calf serum, 100 U/mL penicillin and 30 mg/mL streptomycin sulphate. Finally T.vaginalis culture was harvested in late log phase and centrifuged at 2000 g for 15 min and pellets were stored at −70°C and isolates were confirmed as T.vaginalis by PCR ( 15 , 16 ).…”

Section: Methodsmentioning

confidence: 99%

“…T. vaginalis infection by TVVs could modulate the pathogenicity of T. vaginalis infections by expression of a highly immunogenic T. vaginalis protein, P270 ( 13 ) and also the expression of cysteine proteinases that are assumed to be virulence factors ( 14 ). Genotype of T. vaginalis infected with TVV-1 viruses was evaluated by PCR-RFLP in a study ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

The First Detection of Co-Infection of Double-Stranded RNA Virus 1, 2 and 3 in Iranian Isolates of Trichomonas vaginalis

Bokharaei‐Salim

Esteghamati

Khanaliha

et al. 2020

IJPA

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Background: The Totiviridae family includes a number of double-stranded RNA viruses that can infect Trichomonas vaginalis. Some T. vaginalis isolates are infected with one or more double-stranded RNA (dsRNA) viruses. In this study, different strains of double-stranded RNA virus in Iranian isolates of T. vaginalis were evaluated for the first time in Iran. Methods: Vaginal swabs were collected from 1550 participants who were referred to hospitals associated with Iran University of Medical Sciences, Tehran, Iran from June to November 2018. T. vaginalis isolates were cultured in Diamond's modified medium. After the extraction of nucleic acids using a DNA/RNA extraction kit, RT-PCR was performed and PCR products were purified and sequenced. Results: In general 9 (0.6%) isolates were confirmed as T. vaginalis among 1550 collected vaginal samples. Among 9 isolates of T. vaginalis, three of them were infected with TVV1. One isolate has multiple infections with T. vaginalis virus (TVV1, TVV2 and TVV3) as coinfection. The nucleotide BLAST indicated that the T. vaginalis virus 1(TVV1) isolates were most closely related to TVV1-OC5, TVV1-UR1-1.The T. vaginalis virus 2 (TVV2) sequence had also a similarity with TVV2-UR1-1, TVV2-UR1 and TVV2-OC3. The sequence of T. vaginalis virus 3(TVV3) had similarity with TVV3-OC5, TVV3-UR1-1 and TVV3-UR1. Conclusion: Three dsRNA viruses T. vaginalis virus (TVV1, TVV2 and TVV3) were detected using RT-PCR in T. vaginalis Iranian isolates. The coinfection of TVV1, TVV2 and TVV3 in one isolate of T.vaginalis was observed for the first time in Iran.

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“…gallinae was reported in Australia [32] , and Europe [33] . Utilizing PCR-RFLP targeting actin gene, three studies were carried out in Turkey [34] , China [31} and Iran [35] . Among twenty T. vaginalis isolates from symptomatic females, genotype E was the most common, followed by G, with only one isolate assigned for H, and two mixed isolates of genotypes E and H [34] .…”

mentioning

confidence: 99%

“…In another report, out of 68 T. vaginalis isolates, two E and H genotypes were found in addition to mixed genotypes in three isolates [31] . Recently, investigators detected three genotypes (E, G and I) in Iranian isolates of T. vaginalis infected with dsRNA viruses [35] . Moreover, the majority of isolates obtained from HIV positive women were parasites from genotype II [28] .…”

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confidence: 99%

Genotyping of Trichomonas vaginalis isolates from Egypt

Mohamed

Elleboudy

Hussein

et al. 2019

Parasitologists United Journal

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Trichomoniasis is the most prevalent non-viral sexually transmitted infection worldwide. It has been estimated that 276 million new infections occurred in 2008 with 11.5% increase over the 2005 incidence rate [1]. Worldwide, the estimated prevalence rates vary with geographic location, age, race, community and the method used for diagnosis. In 2016, the WHO global estimate for trichomoniasis in women was estimated at 5.3% and 0.6% in men with a total incidence of 156 million cases [2]. In Egypt, a prevalence rate of 8.0% was recently documented among symptomatic women in Beni Suef Governorate [3] ; in addition to 11% in Benha, Qalyubia Governorate [4] , and 5% in Cairo [5]. Another older study recorded prevalence rate of 27-57% using different techniques for diagnosing trichomoniasis in Shebin El-Kom, Menoufia Governorate [6]. While infections are mostly asymptomatic in men, women usually present with vaginal discharge, pruritus and dysuria [7,8]. Trichomoniasis is considered an important risk factor for herpes simplex virus type II infections and HIV transmission and acquisition [9,10]. Data from studies in Africa recorded an increase in HIV transmission associated with trichomoniasis [11-13]. In an American study, the investigators detected T. vaginalis in 20% of HIV-infected pregnant women using PCR assay [14]. Moreover, it may be associated with cervical cytological abnormalities and cervical cancer [15-18]. Such a difference in clinical traits as virulence, pathogenicity and drug resistance, signifies the need to link phenotypic variation to genotype [19]. In Egypt, former attempts to investigate the diversity of T. vaginalis included isoenzyme patterns [20] , serotyping [21] , immunoblotting [22,23] , biological variability [24,25] and HSP70-RFLP [26]. These studies concluded that the different clinical isolates have different and common patterns at the levels of antigens, immunogens, pathogenicity and MTZ resistance. Molecular typing methods revealed a two-type population structure for T. vaginalis, type I and type II [27-31]. Genotype I infections were found to have lower probability of associated discolored discharge, especially bloody discharge, bleeding during physical examination, or presence of greater than 20% clue cells. Infections with genotype II were significantly associated with these pathological findings. The increase in clue cells seen with genotype II are indicative of bacterial vaginosis, providing a change in the vaginal microenvironment that is favorable for infection with other microbial pathogens. Genetic diversity for T.

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Prevalence and Genotyping of Trichomonas vaginalis Infected to dsRNA Virus by PCR–Restriction Fragment Length Poly-morphism (RFLP)

Cited by 4 publications

References 29 publications

Establishment and application of a CRISPR-Cas12a-based RPA-LFS and fluorescence for the detection of Trichomonas vaginalis

Establishment and application of a CRISPR-Cas12a-based RPA-LFS and fluorescence for the detection of Trichomonas vaginalis

The First Detection of Co-Infection of Double-Stranded RNA Virus 1, 2 and 3 in Iranian Isolates of Trichomonas vaginalis

Genotyping of Trichomonas vaginalis isolates from Egypt

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