MOLECULAR TYPING OF ENDEMIC ISOLATES OF LEPTOSPIRES IN GUILANM,IRAN BY RESTRICTION ENDONUCLEASE ANALYSIS (REA)

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ASMAR M.; HONARMAND H.R.; FATALAHI M.

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Journal: JOURNAL OF BIOLOGY SCIENCE Year:2008 | Volume:2 | Issue:2 (SERIES NO. 5) Page(s): 1-12

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Title

MOLECULAR TYPING OF ENDEMIC ISOLATES OF LEPTOSPIRES IN GUILANM,IRAN BY RESTRICTION ENDONUCLEASE ANALYSIS (REA)

Author(s)

ASMAR M. | HONARMAND H.R. | FATALAHI M. | Issue Writer Certificate

Keywords

GUILANQ3

LEPTOSPIRAQ3

MOLECULARTYPINGQ3

SEROGROUPQ3

Abstract

Leptospirisis is an important zoonosis disease with most widespread in the world. It is more common in regions with warm or temperate and humid climate and with high load of domestic and wild animals, like flat area of GUILAN province, northern of Iran, and with all situation favor to prevalence of leptospirosis. Lots of domestic and and wild animals are source of pathogenic leptospires and also all surface waters are source of saprophytic leptospires and transit medium of nonpathogenic leptospires. Seven pathogenic including more than 200 serovar in 23 SEROGROUP and also 3 nonpathogenic species with several SEROGROUP and serovars are classified until now. Isolation and characterization of endemic strains is a basic step in revealing epidemiological features of the disease and in detecting source(s) of leptospirosis in any endemic area. In this study we typed several strains which we isolated from surface waters of the central region flat area of GUILAN province, which is the only endemic area of human leptospirosis in the country. 301 samples were taken from rice paddies of different parts of the area in spring and summer of 2006. Each sample was filterd by syringeal 22 nm filter and cultured in liquid EMJh medium contained 200 mg/ml 5-flourouracil, checked by dark field microscope every week, until 3 month. Positive samples were cultured in high volume (300 ml), then centrifuge in 13000rpm for 20 min and two times washing (by PBS, pH=7.2), Dna extracted by using Phenol-Chloroform method. Then DNA concentration was measured by using photometer in 260nm.For each positive samolea,3 mg DNa was mixed with 3 ml REA buffer, 9ml sterile ddH2o,and 2ml REA enzyme (ecorI, and HindIII), and incubated in 37°c for 3 hours. Then, after adding 3ml loading buffer, electrophrosis was done in 1%gel agarose, with70 volt for one hour. Band profile of each positive sample was compared with band profiles of representative serovars of all SEROGROUPs to detect its identity. 168 of 301 samples were positive in first round of culturing, but only 33 samples became high rate bacterial numbers in the following culturing stage and were in good position to perform ERA. Band profiles of these samples were similar to some pathogenic (Interrogans, Kirchnery, and Borgpetersenii), and saprophytic (Biflexa, and Wolbachia) species, belong to pathogenic SEROGROUPs: Icterohaemorrahgia, Pomona, Ballum, Hardjo, kanicola, and grippotyphosa, and finally nonpathogenic SEROGROUPs: Andamana, Patoc, and semaranga. Preparing a database of band profiles of main serovars of all SEROGROUPs by REA method, which is a more simple molecular typing way, it is possible to characterize field isolates, mainly in specie and SEROGROUP level. Our study showes that there are some pathogenic serovars mixed to saprophytic ones, in rice paddies, so surface waters can be an important source of pathogenic strains.

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بررسی شیوع لپتوسپیروز در استان گیلان در سال 1383

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APA: Copy

ASMAR, M., HONARMAND, H.R., & FATALAHI, M.. (2008). MOLECULAR TYPING OF ENDEMIC ISOLATES OF LEPTOSPIRES IN GUILANM,IRAN BY RESTRICTION ENDONUCLEASE ANALYSIS (REA). JOURNAL OF BIOLOGY SCIENCE, 2(2 (SERIES NO. 5)), 1-12. SID. https://sid.ir/paper/164237/en

Vancouver: Copy

ASMAR M., HONARMAND H.R., FATALAHI M.. MOLECULAR TYPING OF ENDEMIC ISOLATES OF LEPTOSPIRES IN GUILANM,IRAN BY RESTRICTION ENDONUCLEASE ANALYSIS (REA). JOURNAL OF BIOLOGY SCIENCE[Internet]. 2008;2(2 (SERIES NO. 5)):1-12. Available from: https://sid.ir/paper/164237/en

IEEE: Copy

M. ASMAR, H.R. HONARMAND, and M. FATALAHI, “MOLECULAR TYPING OF ENDEMIC ISOLATES OF LEPTOSPIRES IN GUILANM,IRAN BY RESTRICTION ENDONUCLEASE ANALYSIS (REA),” JOURNAL OF BIOLOGY SCIENCE, vol. 2, no. 2 (SERIES NO. 5), pp. 1–12, 2008, [Online]. Available: https://sid.ir/paper/164237/en

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