ABSTRACT
Objective: To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA. Methods: The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions of T. gondii DNA ranging from 1 ng to 0.01 fg. DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays. Results: After testing LAMP and nested-PCR in duplicate, the detection limit of RE-LAMP, B1-LAMP, RE-nested PCR and B1-nested PCR assays was one fg, 100 fg, 1 pg and 10 pg of T. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection of T. gondii DNA. Conclusions: The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T. gondii. Furthermore, these findings indicate that primers based on the RE are more suitable than those based on the B1 gene. However, the B1-LAMP assay has potential as a diagnostic tool for detection of T. gondii.
ABSTRACT
OBJECTIVE@#To compare analytical sensitivity and specificity of a newly described DNA amplification technique, LAMP and nested PCR assay targeting the RE and B1 genes for the detection of Toxoplasma gondii (T. gondii) DNA.@*METHODS@#The analytical sensitivity of LAMP and nested-PCR was obtained against10-fold serial dilutions of T. gondii DNA ranging from 1 ng to 0.01 fg. DNA samples of other parasites and human chromosomal DNA were used to determine the specificity of molecular assays.@*RESULTS@#After testing LAMP and nested-PCR in duplicate, the detection limit of RE-LAMP, B1-LAMP, RE-nested PCR and B1-nested PCR assays was one fg, 100 fg, 1 pg and 10 pg of T. gondii DNA respectively. All the LAMP assays and nested PCRs were 100% specific. The RE-LAMP assay revealed the most sensitivity for the detection of T. gondii DNA.@*CONCLUSIONS@#The obtained results demonstrate that the LAMP technique has a greater sensitivity for detection of T. gondii. Furthermore, these findings indicate that primers based on the RE are more suitable than those based on the B1 gene. However, the B1-LAMP assay has potential as a diagnostic tool for detection of T. gondii.
ABSTRACT
Background: Blastocystis is a zoonotic protozoan parasite habit in intestinal tract of humans and wide range of animals. Because of the mysterious nature and unknown or less-known aspects of Blastocystis, comprehensive information about epidemiology of this parasite is not available. The objective of this study was to investigate the available parasitology studies during the last decade in Iran and determine the prevalence of Blastocystis and its position among other intestinal parasites. As well as, investigate the effective factors in its prevalence.
Materials and Methods: All available studies related to the prevalence of intestinal parasites in Iran during the recent decade were collected using information databases. After determinant the mean prevalence of each parasite, the most common parasites, and effective factors on their prevalence were assessed and analyzed.
Results: Different studies showed that the most common intestinal parasite at this period of time was Blastocystis spp. with 14.6% prevalence rate. Additionally, in 44.5% of cases Blastocystis spp. allocated the first and in 100% of cases, the first to third rank of the most common intestinal parasites in Iran. Giardia lamblia and Entamoeba coli were in the next category.
Conclusion: To our knowledge, the present study is the first survey in which the Blastocystis spp. introduces as one of the most common intestinal parasites in human. Various factors, including the low sensitivity of routine diagnostic methods, hosts multiplicity, easy transportation and unknown impressive factors are effective in the increased prevalence of this parasite. The results of this study could improve the attitude of teachers and researchers towards Blastocystis spp.
ABSTRACT
Entamoeba moshkovskii and E. dispar are impossible to differentiate microscopically from the pathogenic species E. histolytica. Multiplex polymerase chain reaction [Multiplex PCR] is a widespread molecular biology technique for amplification of multiple targets in a single PCR experiment. For detection and differentiation of the three-microscopy indistinguishable Entamoeba species in human, multiplex PCR assay using different DNA extraction methods was studied. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. A 166-bp PCR product with E. histolytica DNA, a 580-bp product with E. moshkovskii DNA and a 752-bp product with E. dispar DNA were generated in a single-round and multiplex PCR reaction. We recommend this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys