Molecular and Serological Detection of Acute and Latent Toxoplasmosis Using Real-Time PCR and ELISA Techniques in Blood Donors of Rafsanjan City, Iran, 2013
Background: The differentiation between acute and latent forms of the Toxoplasma gondii (T. gondii) infection is still considered as a complicated issue. This study was aimed to elucidate the status of infection in the blood donors and the probable importance of blood transfusion in the transmission of the infection through detecting both immunological and genetic markers of acute and latent infection.
Methods: Totally 235 blood samples from blood donors were collected. The levels of anti-T. gondii IgG and IgM antibodies were examined by specific ELISA kits. cDNA were synthesized from total extracted mRNA molecules from the serum samples and SAG1 gene, specific for tachyzoite form, were amplified using Real-Time PCR technique.Demographic information of study subjects including their gender, age, job,and habitat were recorded.
Results: Out of 235 serum samples, 80 (34.04%) and 4 (1.71%) were positive regarding anti-T. gondii IgG and IgM antibodies, respectively. Real-Time PCR results showed that 14 out of 200 (6.97%) of blood donor had mRNA molecules of SAG1 gene. The positive results of Real-Time PCR of SAG1 in female gender and housekeepers were significantly higher than those of male gender and other job categories.
Conclusion: The prevalence of chronic and acute infection is high in Iranian blood donors. Additionally, evaluation of antibodies could not be reliable, because several donors negative for anti-T. gondii IgM antibodies had detectable SAG1 mRNA molecules.Hence, it seems that molecular diagnostic tests are essential to detect acute infections.
Pusch L, Romeike B, Deckert M, Mawrin C. Persistent Toxoplasma bradyzoite cysts in the brain: incidental finding in an immunocompetent patient without evidence of a toxoplasmosis. Clin Neuropathol. 2009; 28(3):210-2.
Di Cristina M, Del Porto P, Buffolano W, Beghetto E, Spadoni A, Guglietta S, et al. The Toxoplasma gondii bradyzoite antigens BAG1 and MAG1 induce early humoral and cellmediated immune responses upon human infection. Microbes Infect. 2004; 6(2):164-71.
Flegr J, Havlicek J, Kodym P, Maly M, Smahel Z. Increased risk of traffic accidents in subjects with latent toxoplasmosis: a retrospective casecontrol study. BMC Infect Dis. 2002; 2:11.
Ahmad D, Mehdi S, Sayed HH, Sayed AK, Shirzad G. Serological survey of Toxoplasma gondii in schizophrenia patients referred to Psychiatric Hospital, Sari City, Iran. Trop Biomed. 2010; 27(3):476-82.
Fallah M, Rabiee S, Matini M, Taherkhani H. Seroepidemiology of toxoplasmosis in primigravida women in Hamadan, Islamic Republic of Iran, 2004. East Mediterr Health J. 2008; 14(1):163-71.
Hashemi HJ, Saraei M. Seroprevalence of Toxoplasma gondii in unmarried women in Qazvin, Islamic Republic of Iran. East Mediterr Health J. 2010; 16(1):24-8.
Keshavarz H, Nateghpour M, Eskandari S. A Seroepidemiologic Survey Of Toxoplasmosis In Islamshahr District Of Tehran, Iran. Modares Journal of Medical Sciences (Pathobiology). 2004; 6(2):111-9.
Saeedi M, Veghari G, Marjani A. Seroepidemiologic evaluation of anti-Toxoplasma antibodies among women in north of Iran. Pak J Biol Sci. 2007; 10(14):2359-62.
Sharif M, Daryani A, Barzegar G, Nasrolahei M. A seroepidemiological survey for toxoplasmosis among schoolchildren of Sari, Northern Iran. Trop Biomed. 2010; 27(2):220-5.
Sharif M, Ziaei H, Daryani A, Ajami A. Seroepidemiological study of toxoplasmosis in intellectual disability children in rehabilitation centers of northern Iran. Res Dev Disabil. 2007; 28(3):219-24.
Ali Mohammadi H, Fouladi N, Amani F, Safarzadeh M, Pourfarzi F, Mazaheri E. Sero Epidemiological Toxoplasmosis In Pre Marriage Women On The Basis Of Remarriage Tests 2007. Journal of Ardabil University of Medical Sciences (Jaums). 2009; 8(4):408-13.
Keshavarz Valian H, Horri H. Seoprevalence of toxoplasmosis in pergnant women in Kerman City. J Med Council Iran. 1995; 4:322-328.
Keshavarz Valian H, Zare Ranjbar M. Toxoplasmosis in pregnant women and its transmission to infant in Rafsanjan County. Journal of Gilan University of Medical Sciences. 1993; 2(6 &7):28-34.
Montoya J, Boothroyd J, Kovacs J. Section H - Protozoal Diseases, Chapter 279: Toxoplasma gondii. Mandell, Douglas & Bennett's Principles and Practice of Infectious Diseases. 7th ed: Churchill Livingstone, An Imprint of Elsevier; 2009.
Asgari Q, Sarnevesht J, Kalantari M, Sadat SJ, Motazedian MH, Sarkari B. Molecular survey of Toxoplasma infection in sheep and goat from Fars province, Southern Iran. Trop Anim Health Prod. 2011; 43(2):389-92.
Vasina SG, Dunaeva ZV. [On the length of survival of Toxoplasma outside the host organism]. Med Prom SSSR. 1960; 29:451-4.
Siegel SE, Lunde MN, Gelderman AH, Halterman RH, Brown JA, Levine AS, et al. Transmission of toxoplasmosis by leukocyte transfusion. Blood. 1971; 37(4):388-94.
Sundar P, Mahadevan A, Jayshree RS, Subbakrishna DK, Shankar SK. Toxoplasma seroprevalence in healthy voluntary blood donors from urban Karnataka. Indian J Med Res. 2007; 126(1):50-5.
Svobodova V, Literak I. Prevalence of IgM and IgG antibodies to Toxoplasma gondii in blood donors in the Czech Republic. Eur J Epidemiol. 1998; 14(8):803-5.
Alvarado-Esquivel C, Mercado-Suarez MF, Rodriguez-Briones A, Fallad-Torres L, Ayala-Ayala JO, Nevarez-Piedra LJ, et al. Seroepidemiology of infection with Toxoplasma gondii in healthy blood donors of Durango, Mexico. BMC Infect Dis. 2007; 7:75.
Ormazdi H, Sanikhani N, Hadighi R, Akhlaghi L, Memar A, E R. Investigation of antibodies (IgG And IgM) against Toxoplasma Gondii In blood donors referred to Tehran Blood Transfusion Organization by Elisa. Urmia Medical Journal. 2010; 2(21):212-6.
Contini C. Clinical and diagnostic management of toxoplasmosis in the immunocompromised patient. Parassitologia. 2008; 50(1-2):45-50.
Gross U, Holpert M, Goebel S. Impact of stage differentiation on diagnosis of toxoplasmosis. Ann Ist Super Sanita. 2004; 40(1): 65-70.
Press C, Montoya JG, Remington JS. Use of a single serum sample for diagnosis of acute toxoplasmosis in pregnant women and other adults. J Clin Microbiol. 2005; 43(7):3481-3.
Lyons RE, McLeod R, Roberts CW. Toxoplasma gondii tachyzoite-bradyzoite interconversion. Trends Parasitol. 2002; 18(5): 198-201.
Su C, Shwab EK, Zhou P, Zhu XQ, Dubey JP. Moving towards an integrated approach tom molecular detection and identification of Toxoplasma gondii. Parasitology. 2010; 137(1):1-11.
Contini C, Giuliodori M, Cultrera R, Seraceni S. Detection of clinical-stage specific molecular Toxoplasma gondii gene patterns in patients with toxoplasmic lymphadenitis. J Med Microbiol. 2006; 55(Pt 6):771-4.
Mahittikorn A, Wickert H, Sukthana Y. mToxoplasma gondii: Simple duplex RT-PCR assay for detecting SAG1 and BAG1 genes duringm stage conversion in immunosuppressed mice. Exp Parasitol. 2010; 124(2):225-31.
Sajadi SMA, Mirzaei V, Hassanshahi G, Khorramdelazad H, Daredor HY, Hosseini SMH, et al. Decreased Expressions of Toll- Like Receptor 9 and Its Signaling Molecules in Chronic Hepatitis B Virus-Infected Patients. Arch Pathol Lab Med. 2013; 137(11):1674-9.
Hohlfeld P, Daffos F, Costa JM, Thulliez P, Forestier F, Vidaud M. Prenatal diagnosis of congenital toxoplasmosis with a polymerasechain- reaction test on amniotic fluid. N Engl J Med. 1994; 331(11):695-9.
Montoya JG, Parmley S, Liesenfeld O, Jaffe GJ, Remington JS. Use of the polymerase chain reaction for diagnosis of ocular toxoplasmosis. Ophthalmology. 1999; 106(8): 1554-63.
Gianotti N, Cinque P, Castagna A, Novati R, Moro M, Lazzarin A. Diagnosis of toxoplasmic encephalitis in HIV-infected patients. AIDS. 1997; 11(12):1529-30.
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