ß-Globin gene cluster haplotypes of Hb D-Los Angeles in Mazandaran Province, Iran.

Several types of hemoglobin D (Hb D) are distinguishable by DNA analysis, and the aim of this study was to identify the types of Hb D variant and ß-globin gene haplotypes linked to Hb D in Mazandaran Province, northern Iran. Fifty five individuals were identified as Hb D carriers, and PCR-RFLP analysis revealed that all 55 had the Hb D-Los Angeles type. To identify haplotypes associated with the ß(D) allele, family linkage analysis, using the PCR-RFLP method for seven polymorphisms in the ß-globin gene cluster, was carried out on families of 23 of these 55 individuals. We observed three different haplotypes in association with Hb D-Los Angeles. In most cases (91.4%) ß(D) alleles were linked to haplotype I [+ - - - - + +]. Haplotype II [- + + - + + +] and an atypical haplotype [- + + - - + -] were each in association with the ß(D) allele in only one case (4.3%). This is the first report worldwide of the [- + + - - + -] haplotype in association with Hb D-Los Angeles. We conclude that more than 90% of the evaluated Hb D-Los Angeles alleles in Mazandaran have the same origin, and the two rare haplotypes may represent different genetic origins and/or other molecular events, such as gene conversion or recombination, in the region.

Evaluation of gene mutations involved in drug resistance in mycobacterium tuberculosis strains derived from tuberculosis patients in mazandaran, iran, 2013.

Drug resistance (especially multiple drug resistance) in Mycobacterium tuberculosis makes global concerns in treatment and control of tuberculosis. Rapid diagnosis of drug resistant strains of the bacteria has vital importance in the prognosis of the disease. The aim of this study was to identify the mutations responsible for drug resistance in Mycobacterium tuberculosis strains derived from patients with tuberculosis using line probe assay (LPA) method which rapidly detect drug resistant strains and respective mutations. Sputum samples from tuberculosis patients were collected and cultured on Lowenstein- Jensen medium, and then the colonies of Mycobacterium tuberculosis from cultures of 54 bacterial positive cases were randomly chosen for DNA extraction. Bacterial DNA was extracted using standard Cetyl Trimethyl Ammonium Bromide (CTAB) method. In order to identify drug resistant strains and related mutations, LPA method was applied. Three subjects out of 54 investigated cases were resistant to quinolone (5.5%), and resistance to kanamycin/ amikacin, streptomycin, rifampin, and isoniazid were observed in 3 (5.5%), 4 (7.4%), 3 (5.5%), and 2 (3.7%) of the Mycobacterium tuberculosis strains, respectively. In the present study, 4 cases (7.4%) were detected to be resistant to more than one drug. Since LPA is a rapid method that simultaneously detects mutations involved in drug resistance, applying this method in the prediction of drug resistance and selecting appropriate treatment in tuberculosis patients is recommended.

Why does the Iranian national program of screening newborns for G6PD enzyme deficiency miss a large number of affected infants?

PURPOSE: G6PD enzyme deficiency is one of the most prevalent genetic disorders worldwide and it has high incidence rate in Northern provinces of Iran. It was observed that national neonatal screening for G6PD enzyme deficiency fails to detect all affected infants. In order to clarify the cause, this study has been done in Thalassemia Research Center, Sari, Iran. MATERIALS AND METHODS: This was a diagnostic study. The newborns with parents of Mazandarani origin were enrolled. Cord blood from the placental side was collected and used for decolorization test, quantitative enzyme assay (QEA) and DNA study. A heel-prick sample collected on day 3-5 after birth was used for fluorescent spot test (FST). In male cases, QEA was considered as the gold standard. For females, DNA study was considered as the gold standard. Based on QEA test results, neonates with <20% and 20-60% of mean normal enzyme activity were considered as total deficient and partial deficient, respectively. RESULTS: A total of 365 neonates (52.3% females and 47.7% males) were studied. According to FST, 13 male newborns had G6PD deficiency. No deficient female was detected. Decolorization test diagnosed 18 male and one female as G6PD deficient newborns. QEA diagnosed 19 males and 28 females with G6PD enzyme deficiency (26 partial, 2 total deficient cases). DNA analysis detected 14 males as hemizygote and 34 females as heterozygote. CONCLUSION: FST does not have the required sensitivity for newborn screening and QEA is recommended as the preferred method.

Alpha thalassemia gene mutations in neonates from Mazandaran, Iran, 2012.

AIM: Alpha thalassemia is one of the most prevalent disorders worldwide and carrier frequency of the disease is varied in different parts of the world. Although different studies in Iran and Mazandaran province have been carried out to identify different mutations of alpha globin gene among people with low hematological indices, frequencies of these mutations were unknown in general population, and thus the aim of this study was to evaluate the carrier frequencies of alpha globin gene mutations among neonates in Mazandaran. MATERIAL AND METHODS: Four hundred and twelve neonates were collected from a delivery ward of a hospital in Sari. DNA was extracted from their cord blood samples using phenol-chloroform-based method. For the detection of five common alpha thalassemia gene mutations, multiplex-GAP-PCR and PCR-RFLP methods were applied. RESULTS: Sixty three (15.29%, confidence interval, CI 95%: 11.81-18.77) of investigated neonates had at least one of the five evaluated mutations. The -α(3.7) deletion had the highest frequency (9.7%, CI 95%: 6.84-12.56) and none of the neonates had -(Med) double gene deletion. The -α(4.2) deletion, ααα(anti3.7) triplication, and α(-5nt) mutations had frequencies of 4.1% (CI 95%: 2.19-36.01), 2.2% (CI 95%: 0.78-3.62), and 0.49% (CI 95%: -0.18-1.16), respectively. DISCUSSION: Our study showed that in most of the alpha thalassemia carriers just one copy of alpha globin gene was absent and they are not at risk of having children with Hb H disease or hydrops fetalis; however, up to 2.2% of neonates were carriers for ααα(anti3.7) triplication and they will be at risk for having a child with thalassemia intermediate if they marry a person which is a carrier of beta thalassemia.

Is there any relationship between Le(b) antigen expression and Helicobacter pylori infection?

INTRODUCTION: Helicobacter pylori infection is one of the main causes of peptic ulcer. There are some blood groups acting as receptors for the pathogen. Based on this view and previous attempts, we tried to examine the relationship between Lewis blood group and H. pylori infection. MATERIALS AND METHOD: Blood and saliva samples were collected from 60 patients with established peptic ulcer induced by H. pylori. Secretory status of each patient was determined by both direct agglutination and saliva tests. RESULTS: Seventy-two percent of the patients were secretor and expressed Lewis B antigen. This rate in control group was 61%. Statistical analysis showed no significant difference between the two groups. CONCLUSION: This study did not find any correlation between Le(b) antigen expression and presence of H. pylori-induced peptic ulcer. It is now recommended that other factors like Lewis(x) and sialyl Lewis(x) should be investigated in binding, colonization and virulence of H. pylori infection in the future.

Detection of Rare Beta Globin Gene Mutation [+22 5UTR(G>A)] in an Infant, Despite Prenatal Screening.

Background. Beta thalassemia is one of the most common hereditary disorders worldwide. In Iran, it is frequently reported from northern and southern provinces. In order to prevent child birth to be affected by this complication, prenatal screening and diagnosis are carried out nationwide. However, in some instances, this program is unable to identify rare mutations leading to thalassemia. Case Presentation. A married couple, who took part in prenatal screening and diagnosis, gave birth to a child who is affected by thalassemia major. After several molecular examinations, a rare mutation [+22 5UTR (G>A)] in compound heterozygote state along with a common mutation [codon 8 (-AA)] was found. Conclusion. This case study suggests that more advanced molecular evaluations must be integrated in prenatal screening programs to identify rare mutations and antenatal diagnosis of thalassemia cases.

Report of haemoglobin J-Toronto and alpha thalassemia in a family from North of Iran.

We report of an Iranian family with history of a rare haemoglobin variant, Haemoglobin J associated with alpha thalassemia, discovered while performing premarital thalassemia screening. In the present study we report the first case of haemoglobin J-Toronto [alpha 5 (A3) Ala > Asp] on -globin gene, found in a 16-year-old female from Mazandaran Province, North of Iran. Further investigation characterized the same mutation for mother and brother of the proband, whilst mother was also a carrier of an alpha thalassemia gene mutation (-alpha3.7). Haemoglobin J-Toronto was previously just reported from Canada and has not been found in any part of Iran.

Hemoglobin Q-Iran detected in family members from Northern Iran: a case report.

INTRODUCTION: Hemoglobin Q-Iran (α75Asp→His) is an important member of the hemoglobin Q family, molecularly characterized by the replacement of aspartic acid by histidine. The first report of hemoglobin Q-Iran and the nomenclature of this hemoglobinopathy dates back to 1970. Iran is known as a country with a high prevalence of α- and ß-thalassemia and different types of hemoglobinopathy. Many of these variants are yet to be identified as the practice of molecular laboratory techniques is limited in this part of the world. Applying such molecular methods, we report the first hemoglobin Q-Iran cases in Northern Iran. CASE PRESENTATION: An unusual band was detected in an isoelectric focusing test and cellulose acetate electrophoresis of a sample from a 22-year-old Iranian man from Mazandaran Province. Capillary zone electrophoresis analysis identified this band as hemoglobin Q. A similar band was also detected in his mother's electrophoresis (38 years, Iranian ethnicity). The cases underwent molecular investigation and the presence of a hemoglobin Q-Iran mutation was confirmed by the amplification refractory mutation system polymerase chain reaction method. Direct conventional sequencing revealed a single guanine to cytosine missense mutation (c.226G > C; GAC >CAC) at codon 75 in the α-globin gene in both cases. CONCLUSION: The wide spectrum and high frequency of nondeletional α-globin mutations in Mazandaran Province is remarkable and seem to differ considerably from what has been found in Mediterranean populations. This short communication reports the first cases of patients with hemoglobin Q found in that region.

Detection of Hb Setif in north Iran and the question of its origin: Iranian or multiethnic?

Hb Setif is a rare type of hemoglobinopathy resulting from an aspartic acid to tyrosine substitution at codon 94 (GAC>TAC) of the α2-globin gene. In manual and automated hemoglobin (Hb) electrophoresis examination of the case, an unusual band was detected and the result of subsequent capillary electrophoresis suggested that to be Hb Setif. Carrying out polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing, a typical Hb Setif mutation (GAC>TAC) was identified. The haplotype of the α cluster was + + - M PZ + - - - -. This is the first report of such a hemoglobinopathy in North Iran. Various reports of such Hb variants in Iran and countries in the Mediterranean region and North Africa, suggest that the mutation may have occurred around 6,000 years ago, prior to colonization of Aryans on the Iranian plateau.

Raised CA125 serum level in tubercular peritonitis.

A 24-year old woman was admitted with a history of fever and pelvic pain. ESR and CA125 serum level were high and PPD test was negative. Ultra sound and CT evaluation detected free fluid in abdominopelvic cavity. Laparotomy showed fibrinous strands adhering to and fibrotic sac surrounding the components of abdominal cavity. Opening the sac, grey miliary nodules were spotted and pathologic examination revealed multiple granulomatous lesions. Diagnosed with TBP, patient underwent specific antibiotic therapy and her condition improved significantly following treatment. As laboratory findings and image analysis may be misleading in diagnosis of TBP, diagnostic approach of laparotomy and subsequent pathologic examination is of vital value--particularly in premenopausal female patients to preserve fertility.