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Background: Egypt is a developing country with high population density and a high proportion of people with moderate or low socioeconomic levels. The threat of sexually transmitted diseases, such as trichomoniasis, forms an important public health concern. Trichomoniasis, especially in asymptomatic individuals, remains neglected and poorly addressed .Trichomonas Vaginalis, a parasitic protozoa that causes the sexually transmitted infection trichomoniasis, is the sexually transmitted infection with the largest annual incidence and considered as one of the neglected tropical parasitic infections. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, drug susceptibility and epidemiology of the infection, we conducted analysis of the actin gene of T. vaginalis by applying the Restriction fragment length polymorphism (RFLP) and nucleotide sequencing method. Aim and objectives: To investigate the genetic variability of T.Vaginalis in infecting Egyptian women in Upper Egypt. Patient & methods: This cross-sectional research was conducted during the period from 2019 to 2022 on 150 female patients attending the outpatient clinics outpatient clinics of gynecology and obstetrics at Sohag General Hospital with complaints suggestive of trichomoniasis. Vaginal washout samples were collected, and subjected to direct wet mount microscopic examination, and culture on TYM Diamond's medium, then analysis of the actin gene of T. vaginalis by PCR-RFLP( PCR amplification, restriction enzyme digestion) and detection of DNA fragments by electrophoresis in 2.5–3% (w/v) agarose gels Results: By using direct wet-mount microscopic examination, out of 150 collected wash-out vaginal samples, 12 cases (8%) were positive for T. vaginalis trophozoites, and 16 samples (10.6%) were positive for culture , (13) samples applied to PCR-RFLP ,The restriction enzyme HindII digested the amplified products into three DNA fragments of 60, 213, and 827 bp. RsaI produced 5 DNA fragments of 87, 103\ 106, 236, & 568 bp, & MseI produced 3 different fragments of 204, 315, & 581 bp ,and detection of DNA fragments by electrophoresis in agarose gels . Conclusion: According to our study Genotype E of T. vaginilais is the most prevalent in upper Egypt.
Background: Egypt is a developing country with high population density and a high proportion of people with moderate or low socioeconomic levels. The threat of sexually transmitted diseases, such as trichomoniasis, forms an important public health concern. Trichomoniasis, especially in asymptomatic individuals, remains neglected and poorly addressed .Trichomonas Vaginalis, a parasitic protozoa that causes the sexually transmitted infection trichomoniasis, is the sexually transmitted infection with the largest annual incidence and considered as one of the neglected tropical parasitic infections. More understanding about genetic features of the parasite can be helpful in the study of the pathogenesis, drug susceptibility and epidemiology of the infection, we conducted analysis of the actin gene of T. vaginalis by applying the Restriction fragment length polymorphism (RFLP) and nucleotide sequencing method. Aim and objectives: To investigate the genetic variability of T.Vaginalis in infecting Egyptian women in Upper Egypt. Patient & methods: This cross-sectional research was conducted during the period from 2019 to 2022 on 150 female patients attending the outpatient clinics outpatient clinics of gynecology and obstetrics at Sohag General Hospital with complaints suggestive of trichomoniasis. Vaginal washout samples were collected, and subjected to direct wet mount microscopic examination, and culture on TYM Diamond's medium, then analysis of the actin gene of T. vaginalis by PCR-RFLP( PCR amplification, restriction enzyme digestion) and detection of DNA fragments by electrophoresis in 2.5–3% (w/v) agarose gels Results: By using direct wet-mount microscopic examination, out of 150 collected wash-out vaginal samples, 12 cases (8%) were positive for T. vaginalis trophozoites, and 16 samples (10.6%) were positive for culture , (13) samples applied to PCR-RFLP ,The restriction enzyme HindII digested the amplified products into three DNA fragments of 60, 213, and 827 bp. RsaI produced 5 DNA fragments of 87, 103\ 106, 236, & 568 bp, & MseI produced 3 different fragments of 204, 315, & 581 bp ,and detection of DNA fragments by electrophoresis in agarose gels . Conclusion: According to our study Genotype E of T. vaginilais is the most prevalent in upper Egypt.
IntroductionTrichomoniasis remains one of the most significant sexually transmitted disease (STDs) for public health. The disease is caused by parasitic protozoa, Trichomonas vaginalis (T. vaginalis), which is often underestimated in tropical medicine. Despite its public health importance, the epidemiology and molecular characteristics of trichomoniasis in Egypt remains poorly understood, particularly in the southern part of the country (Upper Egypt). This study targeted exploring the genetic variability of T. vaginalis infections in Egyptian women living in Upper Egypt using restriction fragment length polymorphism (RFLP).Patient and techniquesThis cross-sectional study included 150 female patients, who visited the gynaecology and obstetrics outpatient clinics at Sohag General Hospital between 2019 and 2022, exhibiting symptoms of trichomoniasis. Vaginal washout samples were collected from each patient and analyzed using three diagnostic techniques: direct wet mount microscopy, culture on TYM Diamond’s medium, and PCR amplification and Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) targeting the actin gene, which was applied to all 16 samples that tested positive in culture. The PCR-RFLP results were then visualized through agarose gels electrophoresis to detect DNA fragments.ResultsOut of 150 vaginal washout samples, 12 cases (8%) tested positive for T. vaginalis trophozoites via direct wet mount microscopy, while 16 samples (10.6%) were positive in culture. Additionally, PCR-RFLP analysis of the 16 culture-positive samples revealed that 13 samples were confirmed positive using this molecular method. The amplified products were digested with the restriction enzyme Hind II, yielding three DNA fragments of 60, 213, and 827 bp, which were then detected by agarose gel electrophoresis. Digestion with RsaI produced five fragments measuring 87, 103/106, 236, and 568 bp, while MseI digestion resulted in three distinct fragments of 204, 315, and 581 bp.ConclusionThis study provides robust baseline data on the prevalence and microscopic characteristics of T. vaginalis in Upper Egypt, while also presenting, for the first time, molecular detection and genotyping and revealed that genotype E is the only prevalent genotype in the region.
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