• 1.

    Bharti AR, Nally JE, Ricaldi JN, Matthias MA, Diaz MM, Lovett MA, Levett PN, Gilman RH, Willig MR, Gotuzzo E, Vinetz JM, 2003. Leptospirosis a zoonotic disease of global importance. Lancet Infect Dis 3: 757771.

    • Search Google Scholar
    • Export Citation
  • 2.

    Mérien F, Amouriaux P, Perolat P, Baranton G, Saint Girons I, 1992. Polymerase chain reaction for detection of Leptospira spp. in clinical samples. J Clin Microbiol 30: 22192224.

    • Search Google Scholar
    • Export Citation
  • 3.

    Léon A, Pronost S, Tapprest J, Foucher N, Blanchard B, André-Fontaine G, Laugier C, Fortier G, Leclercq R, 2006. Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis. J Vet Diagn Invest 18: 218221.

    • Search Google Scholar
    • Export Citation
  • 4.

    Rafyi A, Magami GH, 1968. Leptospirose ovine et caprine. Arch Inst Razi 20: 2538.

  • 5.

    Adler B, Murphy AM, Locarnini SA, Faine S, 1980. Detection of specific anti- leptospiral immunoglobulines M and G in human serum by solid-phase enzyme-linked immunosorbent assay. J Clin Microbiol 11: 452457.

    • Search Google Scholar
    • Export Citation
  • 6.

    Torten M, Shenberg E, Van der Hoeden J, 1966. The use of immunofluorescence in the diagnosis of leptospirosis by a genus-specific antigen. J Infect Dis 116: 537543.

    • Search Google Scholar
    • Export Citation
  • 7.

    Udomsakdi S, Potha U, 1972. The diagnosis of leptospirosis by fluorescent antibody technique using saprophytic Leptospira as a genus-specific antigen. J Med Assoc Thai 55: 101104.

    • Search Google Scholar
    • Export Citation
  • 8.

    Appassakij H, Silpapojakul K, Wansit R, Woodtayakorn J, 1995. Evaluation of the immunofluorescent antibody test for the diagnosis of human leptospirosis. Am J Trop Med Hyg 52: 340343.

    • Search Google Scholar
    • Export Citation
  • 9.

    Djadid ND, Ganji ZF, Gouya MM, Rezvani M, Zakeri S, 2009. A simple and rapid nested polymerase chain reaction-restriction fragment length polymorphism technique for differentiation of pathogenic and nonpathogenic Leptospira spp. Diagn Microbiol Infect Dis 63: 251256.

    • Search Google Scholar
    • Export Citation
  • 10.

    Mansour-Ghanaei F, Sarshad A, Fallah MS, Pourhabibi A, Pourhabibi K, Yousefi-Mashhoor M, 2005. Leptospirosis in Guilan, a northern province of Iran: assessment of the clinical presentation of 74 cases. Med Sci Monit 11: 219223.

    • Search Google Scholar
    • Export Citation
  • 11.

    Laras K, Cao BV, Bounlu K, Nguyen TK, Olson JG, Thongchanh S, Tran NV, Hoang KL, Punjabi N, Ha BK, Ung SA, Insisiengmay S, Watts DM, Beecham HJ, Corwin AL, 2002. The importance of leptospirosis in Southeast Asia. Am J Trop Med Hyg 67: 278286.

    • Search Google Scholar
    • Export Citation
  • 12.

    Yersin C, Bovet P, Mérien F, Wong T, Panowsky J, Perolat P, 1998. Human leptospirosis in the Seychelles (Indian Ocean): a population-based study. Am J Trop Med Hyg 59: 933940.

    • Search Google Scholar
    • Export Citation
  • 13.

    Padre LP, Watt G, Tuazon ML, Gray MR, Laughlin LW, 1988. A serologic survey of rice-field leptospirosis in central Luzon, Philippines. Southeast Asian J Trop Med Public Health 19: 197199.

    • Search Google Scholar
    • Export Citation
  • 14.

    Ciceroni L, Stepan E, Pinto A, Pizzocaro P, Dettori G, Franzin L, Lupidi R, Mansueto S, Manera A, Ioli A, Marcuccio L, Grillo R, Ciarrocchi S, Cinco M, 2000. Epidemiological trend of human leptospirosis in Italy between 1994 and 1996. Eur J Epidemiol 16: 7986.

    • Search Google Scholar
    • Export Citation
  • 15.

    Levett PN, 2001. Leptospirosis. Clin Microbiol Rev 14: 296326.

  • 16.

    Thai KT, Nga TT, Phuong HL, Giao PT, Hung le Q, Binh TQ, Van Nam N, Hartskeerl RA, de Vries PJ, 2008. Seroepidemiology and serological follow-up of anti-leptospiral IgG in children in southern Vietnam. Acta Trop 106: 128131.

    • Search Google Scholar
    • Export Citation
  • 17.

    Romero EC, Bernardo CC, Yasuda PH, 2003. Human leptospirosis: a twenty-nine-year serological study in São Paulo, Brazil. Rev Inst Med Trop Sao Paulo 45: 245248.

    • Search Google Scholar
    • Export Citation
  • 18.

    Cumberland P, Everard CO, Wheeler JG, Levett PN, 2001. Persistence of anti-leptospiral IgM, IgG and agglutinating antibodies in patients presenting with acute febrile illness in Barbados 1979–1989. Eur J Epidemiol 17: 601608.

    • Search Google Scholar
    • Export Citation
  • 19.

    Silva MV, Camargo ED, Batista L, Vaz AJ, Brandão AP, Nakamura PM, Negrão JM, 1995. Behaviour of specific IgM, IgG and IgA class antibodies in human leptospirosis during the acute phase of the disease and during convalescence. J Trop Med Hyg 98: 268272.

    • Search Google Scholar
    • Export Citation
  • 20.

    Wohl JS, 1996. Canine leptospirosis. Compend Contin Educ Pract Vet 18: 12151241.

  • 21.

    Sasaki DM, 2002. Questions stated prevalence of leptospirosis in dogs. J Am Vet Med Assoc 220: 14521453.

  • 22.

    Aslantas O, Ozdemir V, Kiliç S, Babür C, 2005. Seroepidemiology of leptospirosis, toxoplasmosis, and leishmaniosis among dogs in Ankara, Turkey. Vet Parasitol 129: 187191.

    • Search Google Scholar
    • Export Citation
 
 
 
 

 

 
 

 

 

 

 

 

 

Molecular Epidemiology of Leptospirosis in Northern Iran by Nested Polymerase Chain Reaction/Restriction Fragment Length Polymorphism and Sequencing Methods

View More View Less
  • Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Institute Pasteur of Iran, Tehran, Iran; Amol Research Center Branch of Pasteur Institute of Iran, Amole, Mazandaran, Iran

This study was conducted to investigate the prevalence of Leptospira species in Mazandaran Province of Iran by using nested polymerase chain reaction (PCR)/restriction fragment length polymorphism (RFLP) methods and sequencing analysis. Blood samples (n = 119) were collected from humans suspected of having leptospirosis from different parts of the province in 2007. By using an indirect immunofluorescent antibody test (IFAT), we determined that 35 (29.4%) of 119 suspected cases had leptospiral antibody titers ≥ 1:80, which confirmed the diagnosis of leptospirosis. Nested PCR assay also determined that 60 (50.4%) of 119 samples showed Leptospira infection. Furthermore, 44 (73.3%) of 60 confirmed leptospirosis amplified products were subjected to sequencing analysis. Sequence alignment identified L. interrogans, L. kirschneri, and L. wolffii species. All positive cases diagnosed by IFAT or PCR were in patients who reported contact with animals, high-risk occupational activities, and exposure to contaminated water. Therefore, it is important to increase attention about this disease among physicians and to strengthen laboratory capacity for its diagnosis in infected patients in Iran.

Author Notes

*Address correspondence to Sedigheh Zakeri, Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. E-mails: zakeris@yahoo.com or zakeris@pasteur.ac.ir

Financial support: This study was supported by research grant no. 320 from the Pasteur Institute of Iran.

Authors' addresses: Sedigheh Zakeri, Neda Sepahian, Mandana Afsharpad, and Navid D. Djadid, Malaria and Vector Research Group (MVRG), Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. Behzad Esfandiari and Peyman Ziapour, Amol Research Center Branch of Pasteur Institute of Iran, Amole, Mazandaran, Iran.

Reprint requests: Sedigheh Zakeri, Biotechnology Research Centre, Pasteur Institute of Iran, Tehran, Iran, E-mails: zakeris@yahoo.com or zakeris@pasteur.ac.ir.

Save