ABSTRACT
Background and Objective: Iran remains a major stronghold for glanders in the Middle East. In Iran, the non-indigenous Burkholderia mallei Razi 325 strain is used in manufacturing of the mallein, required for malleination of animals. Multi Locus Variable number tandem repeat analysis is currently the standard globally accepted genotyping system for Burkholderia mallei. This study was done to survey the genomic structure of Burkholderia mallei Razi 325, the strain used for industrial production of Mallein
Methods: In this descriptive study, a MLVA genotyping system with 4 previously-characterized loci VNTR140, VNTR1367, VNTR2065, VNTR2971 along with two new loci of VNTR24, VNTR41 was used
Results: Optimization of PCRs resulted in a single protocol that enabled simultaneous amplification of all the six loci. Sequencing of PCR products revealed there were 2, 3, 12, 6, 1 and 2 copies of the unit repeat hold in the genome of the Burkholderia mallei Razi 325 strain. This observation was extended to include the already-whole genome sequenced Chinese Burkholderia mallei ATCC 23344 and Burkholderia mallei BMQ and also Burkholderia mallei SAVP1 strains
Conclusion: The Burkholderia mallei Razi 325 strain is distinguishable from the other three strains through MLVA genotyping method
ABSTRACT
st and ardized genotyping systems in molecular epidemiology of tuberculosis in the world. This sudy was done to determine the Mycobacterium tuberculosis genotyping by MIRU-VNTR method. Methods: This descriptive study was done on sputum, gastric lavage clinical specimens of 53 tuberculosis suspected patients. Fifty-three isolates were identified by 16S rRNAandRv-typing followed by RD typing. They were then subjected to a 12-locus [ETRA, ETRB, ETRC, ETRD, ETRE and ETRF, MIRU-10, MIRU-26, MIRU-39, MIRU-30 plus QUB-11b] MIRU-VNTR typing system. Results: In MIRU-VNTR typing, forty-four types were identified with 13 isolates classified in 4 clustered and the remaining 40 isolates representing 40 orphan patterns. In comparative analysis of MIRU-VNTR loci, MIRU-26 with 7 alleles displayed the highest diversity level [Simpson's diversity index = 0.767. Out of the 53 isolates, only one was identified as Mycobacterium bovis. All the remaining isolates were characterized as Mycobacterium tuberculosis. None of the samples was affected to Mycobacterium complex strain. No evidence of either double or co-infection of the patients with more than one species/strain was detected. Conclusion: While the genomic diversity observed by MIRU-VNTR typing sounds extensive, the population genomic structure on the whole however, seems to be homogenous. Recent transmission between studied patients does not appear to be a frequent event as only 13 isolates representing 4 MIRU-VNTR types, were assumingly epidemic
ABSTRACT
A high level of homogeneity observed within all bacteria in the Mycobacterium tuberculosis complex makes a property that seriously challenges traditional biochemical-based identification methods of these pathogens in the laboratory. The work presented here was conducted to characterize Mycobacterium tuberculosis complex isolates in Golestan, Northern Iran. Between 2008 and 2010, 42 mycobacterial isolates were collected from clinical tuberculosis-suspected patients in Golestan province. The isolates were sub-cultured on fresh Mycobacterium-specific culture media including glycerinated and pyruvated Lowenstein-Jensen slopes. The isolates were subsequently subjected to a PCR-based identification scheme coined Huard-Warren method. This strategy consisted of three individual algorithms namely, 16SrRNA; RV typing [Rv0577, Rv3877.8, Rvl970, Rv3120, Rvl510 and IS 1561] and RD typing [RD1, RD 4, RD9 and RD12]. All isolates were proved to be M. tuberculosis. Furthermore, none of the patients were being infected with any other member of the M. tuberculosis complex or simultaneously co-infected with two mycobacteria. This fundamental observation was independently obtained by specific culture media, RV typing and also RD typing. Considering the fact that cattle and sheep farming play an important role in the economy of the region, absence of Mycobacterium bovis in the studied isolates can be unexpected to some extent. Huard-Warren which is a simple and cost-effective identification method can be used in both reference and regional laboratory for differential diagnosis of tuberculosis
ABSTRACT
Infection with Toxoplasma gondii during pregnancy can lead to severe fetal sequelae. This cross-sectional study aimed to determine T. gondii seropositivity among a sample of young women in Jahrom, Islamic Republic of Iran. Four hundred and three young women with a mean age of 20.3 years who presented for pre-marriage laboratory testing were entered in the study. T. gondii antibodies [IgG and IgM] were measured using ELISA. Of the 403 women, 15% were seropositive for T. gondii antibodies [13% for IgG and 2% for IgM].Seropositivity for T. gondii IgG differed according to age groups but there was no significant difference [P = 0.83]. IgM seropositivity showed the highest rate among women aged < 20 years. Many young women in Jahrom Province are susceptible to primary toxoplasmosis during pregnancy, therefore, appropriate educational programmes to improve knowledge in this population should be implemented to prevent toxoplasmosis-related congenital malformations
Subject(s)
Humans , Female , Toxoplasmosis/prevention & control , Toxoplasma/immunology , Cross-Sectional Studies , Pregnancy Complications/parasitology , Enzyme-Linked Immunosorbent Assay , Health Education , Congenital Abnormalities/parasitologyABSTRACT
Pigeons are extensively kept for homing and racing purposes in Iran. The main objective of this study was to investigate dissemination of M. avium subsp. avium [MAA] in pigeon aviaries in Tabriz, North-western Iran. Postmortem pathologic specimens from thirty-nine out of 140 birds collected from private flocks [n = 3], were subjected to bacterial culture out of which 3-4 mycobacterial isolates were recovered. Applying a five-PCR diagnostic algorithm targeting short but definitive stretches of 16S rRNA and RV0577 genes, IS6110, IS901 and IS1245 genomic loci, proved all the isolates were MAA. They were either IS901+/IS1245+ [n = 22] or IS901+/IS1245- [n = 12]. When four healthy cattle sensitized against Mycobacterium bovis AN5 and Mycobacterium avium D4 were tuberculinated, the results confirmed the observed skin reactions against bovine tuberculin in animals sensitized with M. avium were large enough to complicate test interpretation. We believe the extent of such epidemiological impact deserves further investigation if progress in control of bovine tuberculosis is intended
ABSTRACT
Mycobacterium tuberculosis complex is consisted of homogenous organisms. They are slowly growing mycobacteria and their isolation and identification are difficult and time consuming. Differentiation of Mycobacterium bovis, causative mammalian tuberculosis, from other members of Mycobacterium tuberculosis complex is very important in epidemiology and control of disease in humans and animals. The aim of this study was to evaluate a molecular method to differentiate Mycobacteriom bovis from Mycobacterium tuberculosis. DNA human isolates of Mycobacterium tuberculosis [n=6] and Mycobacterium bovis isolates [50] were extracted and used as template in PCR. A 548bp fragment of oxyR pseudogene was amplified and digested with Alul endo nuclease. The nucleotide 285 could be adenine [M. bovis] or guanine [M. tuberculosis]. Such variation produces different restriction site for Alul. There were three incisive fragments in all Mycobacterium bovis and Mycobacterium bovis BCG strains and one incisive fragment in other members of Mycobacterium tuberculosis complex. PCR-RFLP method on 548bp fragment of oxyR gene is a rapid and accurate method to differentiate Mycobacterium bovis and Mycobacterium bovis BCG from other members of Mycobacterium tuberculosis complex