23
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Comparison of Placenta PCR and Maternal Serology of Aborted Women for Detection of Toxoplasma gondii in Ardabil, Iran

      research-article

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Primary maternal infection with toxoplasmosis during pregnancy is frequently associated with transplacental transmission of the parasite to the fetus. This study was conducted to test the utility of PCR assay to detect recent infections with Toxoplasma in aborted women at various gestational ages who referred to Obstetrics and Gynecology Department of Alavi Hospital in Ardabil during 2014 and 2016. Two hundred women with a history of single or repeated abortion were investigated in this study. Blood samples were tested for specific anti- Toxoplasma IgM and IgG antibodies by ELISA. According to the results, 53.5% of the women under study were positive for anti- Toxoplasma antibodies: 4.0% of them had IgM, 43.0% had IgG, and 6.5% had both IgM and IgG. Subsequently, Nested-PCR analysis was used to detect T. gondii DNA in the placenta of subjects. In 10.5% of the women, the results were positive for 529 bp element of T. gondii. Among them, 5 (23.8%) cases were IgM positive, 1 (4.8%) case was IgG positive, and 11 (52.4%) were both IgM and IgG positive. In 4 (19.0%) patients, none of the antibodies were found to be positive. In total, 16 patients had positive results in both ELISA and PCR methods, and 174 cases had negative results for new infection. The findings of this study revealed that T. gondii might be one of the significant factors leading to abortion, and that the analysis of placenta can be important in order to achieve increased detection sensitivity.

          Related collections

          Most cited references18

          • Record: found
          • Abstract: found
          • Article: not found

          Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR.

          We have identified a novel 529bp fragment that is repeated 200- to 300-fold in the genome of Toxoplasma gondii. This 529bp fragment was utilised for the development of a very sensitive and specific PCR for diagnostic purposes, and a quantitative competitive-PCR for the evaluation of cyst numbers in the brains of chronically infected mice. The 529bp fragment was found in all 60 strains of T. gondii tested, and it discriminates DNA of T. gondii from that of other parasites. Toxoplasma gondii DNA was detected in amniotic fluid of patients, as well as in various tissues from infected mice. Polymerase chain reaction with the 529bp fragment was more sensitive than with the 35-copy B1 gene. For the quantitative competitive-PCR, a 410-bp competitor molecule was co-amplified with similar efficiency as the 529bp fragment. Quantitative competitive-PCR produced a linear relationship between the relative amounts of PCR product and the number of tachyzoites in the range of 10(2)-10(4) tachyzoites and 100-3000 tissue cysts. A highly significant correlation between visual counting of brain cysts and quantitative competitive-PCR was obtained in mice chronically infected with Toxoplasma. Thus, quantitative competitive-PCR with the 529bp fragment can be used as an alternative for the tedious visual counting of brain cysts in experimental animals. With the quantitative competitive-PCR, furthermore, we could confirm the copy number of the 529bp fragment in tachyzoites and estimate the number of bradyzoites per cyst.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Comparison of PCR detection methods for B1, P30, and 18S rDNA genes of T. gondii in aqueous humor.

            Comparison of polymerase chain reaction (PCR) amplification of three Toxoplasma gondii genes in aqueous humor. Nested PCRs carried out using published methods were optimized for maximum sensitivity and specificity. Five pairs of oligonucleotide primers, directed against the B1, P30, and ribosomal genes, were used and compared to determine which sequences were most effective in detecting T. gondii DNA. Methods were developed with DNA templates in water and were subsequently applied to both normal and inflamed aqueous. After one round of PCR amplification, P30 and ribosomal primers were able to detect 1 pg genomic T. gondii DNA. However, those directed against the B1 gene were able to detect 50 fg (approximately single tachyzoite). This level of sensitivity was also achieved using the P30 primers after a second round of PCR; however, only primers based on the B1 gene maintained this level of sensitivity in both normal and inflamed aqueous. B1-specific primers did not amplify sequences from fungal, bacterial, or human lymphocyte DNA. The sensitivity of T. gondii detection using B1 gene-specific primers was not compromised when large amounts of human lymphocyte DNA were present, and application to an ocular sample or retinal section from patients with toxoplasma chorioretinitis was successful in confirming the presence of T. gondii DNA. The B1 PCR protocol appears to be the most sensitive protocol in the detection of T. gondii DNA and has been successful in identification of T. gondii DNA in ocular fluids and retinal sections. This provides direct evidence of the presence of T. gondii within the eye and may therefore help in the management of toxoplasma retinochoroiditis.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: found
              Is Open Access

              Prevalence of toxoplasmosis in pregnant women and vertical transmission of Toxoplasma gondii in patients from basic units of health from Gurupi, Tocantins, Brazil, from 2012 to 2014

              Introduction Toxoplasmosis is a parasitary disease that presents high rates of gestational and congenital infection worldwide being therefore considered a public health problem and a neglected disease. Objective To determine the prevalence of toxoplasmosis amongst pregnant women and vertical transmission of Toxoplasma gondii in their newborns attended in the Basic Units of Health (BUH) from the city of Gurupi, state of Tocantins, Brazil. Methods A prevalence study was performed, including 487 pregnant women and their newborns attended in the BUH of the urban zone of the city of Gurupi, state of Tocantins, Brazil, during the period from February 2012 to February 2014. The selection of the pregnant women occurred by convenience. In the antenatal admission they were invited to participate in this study. Three samples of peripheral blood were collected for the detection of specific anti-T. gondii IgG, IgM and IgA through ELISA, for the polimerase chain reaction (PCR) and IgG avidity during pregnancy. When IgM antibodies were detected the fetal and newborn infection investigation took place. The newborn was investigated right after birth and after one year of age through serology and PCR to confirm/exclude the vertical transmission. The analyses were performed in the Studies of the Host-Parasite Relationship Laboratory (LAERPH, IPTSP-UFG), Goiania, state of Goias, Brazil. The results were inserted in a data bank in Epi-Info 3.3.2 statistic software in which the analysis was performed with p≤5%. Results The toxoplasmosis infection was detected in 68.37% (333/487, CI95%: 64.62–72.86). The toxoplasmosis chronic infection prevalence was of 63.03% (307/487, CI95%: 58.74–67.32). The prevalence of maternal acute infection was of 5.33% (26/487; CI95%: 3.3–7.3) suspected by IgM antibodies detection in the peripheral blood. The prevalence of confirmed vertical transmission was of 28% (7/25; CI95%: 10.4–45.6). Conclusions These results show an elevated prevalence of toxoplasmosis in pregnant women and vertical transmission of T. gondii in the city of Gurupi, state of Tocantins, Brazil.
                Bookmark

                Author and article information

                Journal
                Korean J Parasitol
                Korean J. Parasitol
                The Korean Journal of Parasitology
                The Korean Society for Parasitology and Tropical Medicine
                0023-4001
                1738-0006
                December 2017
                31 December 2017
                : 55
                : 6
                : 607-611
                Affiliations
                [1 ]Department of Internal Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
                [2 ]Department of Parasitology and Mycology, Faculty of Specialized Veterinary Science, Islamic Azad University, Science and Research Branch, Tehran, Iran
                [3 ]Department of Obstetrics and Gynecological, Ardabil University of Medical Sciences, Ardabil, Iran
                [4 ]Ardabil University of Medical Sciences, Ardabil, Iran
                [5 ]Department of Microbiology, Ardabil University of Medical Sciences, Ardabil, Iran
                Author notes
                [* ]Corresponding author ( rzshhbz@ 123456yahoo.com )
                Article
                kjp-55-6-607
                10.3347/kjp.2017.55.6.607
                5776894
                29320815
                9e19998f-f2cb-4ad3-a109-43203d9f0cb8
                Copyright © 2017 by The Korean Society for Parasitology and Tropical Medicine

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 13 July 2017
                : 26 November 2017
                : 27 November 2017
                Categories
                Original Article

                Parasitology
                toxoplasma gondii,abortion,blood,nested-pcr,elisa,iran
                Parasitology
                toxoplasma gondii, abortion, blood, nested-pcr, elisa, iran

                Comments

                Comment on this article