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Diagnosis of Enteroviral-associated-Meningitis by RT-PCR for the First Time in Iraq

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Abstract

Introduction: A protocol for testing cerebrospinal fluid specimens using RT-PCR for the diagnosis of central nervous system infections was developed 2 and used totest 20 CSF specimens.Objectives: A-Evaluation of efficiency of In-house prepared Extraction reagent. BAmplified of Enteroviral-RNA genome by single-step RT-PCR. Methods: 20cerebrospinal fluid specimens were included. RNA was extracted from 250μl by method Chomcynki, 1993. EV-RT-PCR was performed for all specimens studied.Results: We successfully isolated total RNA from 20 CSF specimens of AM cases.Ten of 20 specimens were positive by RT-PCR.Conclusions: Single-step extraction method was allowed isolation of RNA in 4hr.and provided both high yield and purity of un-degraded RNA preparation. EV-RTPCR detection test is suitable for clinical diagnosis of EV-related meningitis andmay improve the management of EV-related neurologic syndromes.

لتشخیص إصابات الجھاز العصبي RT-PCR◌ٌٌ مقدمة : لفحص نماذج سائل النخاع الشوكي, استخدم تقنیةالمركزي وقد تم فحصعشرین نموذج من سائل النخاع الشوكي.الھدف :في استخلاص الحامض النووي المنقوص (TRI-Reagent) أ- تقییم كفاءة الخلیط الثلاثي للموادRNA الأوكسجین.Single step-RT-PCRٌ باستخدام تقنیة Enteroviral-RNA-genome ب - إمكانیة مضاعفةطریقة العمل: تضمن البحث ( 20 ) نموذج سائل النخاع الشوكي (حجم 250 مایكرولیتر من سائل النخاعوحضرت Chomcynki, تبعا لطریقة 1993 RNA الشوكي) واستخدم في استخلاص الحامض النوويلفحصجمیع نماذج الدراسة. EV-RT-PCR تقنیةمن عشرین نموذج سائل نخاع شوكي من حالات RNA النتائج: لقد نجحنا باستخلاص الحامض النووي. EV-RT-PCR عشرة منھا كانت موجبة في فحص ,aseptic meningitis التھاب السحایاخلال أربع RNA الاستنتاج: أظھرت طریقة الاستخلاصذات الخطوة المنفردة قابلیتھا على استخلاصوأعطتنا منتوج جید والحامض النووي (TRI-Reagent) ساعات فقط باستخدام الخلیط الثلاثي للموادفحص ملائم للتشخیص الطبي لالتھاب السحایا EV-RT-PCR المستخلص ذا نقاوة عالیة. یعتبرالمتسبب من الإصابة بالفیروسات المعویة وباستخدامھ یطور ویحسن معالجة المتلازمات العصبیة المتسببةعن الفیروسات المعویة.


Article
دراسة تشخيصية ووبائية لداء الليشمانيا الاحشائية Visceral Leishmaniasis باستعمال تقنية PCR في محافظة ذي قار

Author: فاضل عباس منشد العبادي
Journal: Univesity of Thi-Qar Journal مجلة جامعة ذي قار العلمية ISSN: ISSN: 27066894 (Online) ISSN: 27066908 (Print) Year: 2008 Volume: 4 Issue: 1 Pages: 1-8
Publisher: Thi-Qar University جامعة ذي قار

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Article
Molecular Detection of Some Mutations Associated with Βeta-Thalassaemia in Iraq

Authors: Massoud Houshmand --- Ali A. Al-Zaag --- Norrya A. Ali --- Waleed K. Al-Murrani --- et al.
Journal: Karbala Journal of Medicine مجلة كربلاء الطبية ISSN: 19905483 Year: 2008 Volume: 2 no.4, 5 Issue: 3 Pages: 263-274
Publisher: Kerbala University جامعة كربلاء

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Abstract

the study was carried out in period between “1 August 2005 to 30 of December 2006”.Blood samples of 80 clinically thalassaemic patients were collected from three thalassaemiacenters in Iraq, namely: Ibn Albalady central thalassaemia center in Baghdad, and also fromKerbalaa and Al-Qadissya governorates. Blood samples were also collected from 56 apparentlyhealthy individuals to serve as a control group.DNA was isolated from blood and used for molecular detection of seven types of β-thalassaemiamutations (IVS1 nt.1 G--A , IVS1nt.5 G--C, IVS1 nt.6 T--C, IVS1 nt.110 G --A, codon 39 C--T,IVS2 nt.1G--A, and IVS2 nt.745 C--G) using the PCR based technique called ARMS(Amplification Refractory Mutation System).Five out of seven of these diagnosed mutations were reported for the first time in Iraq, and themost frequent β-thalassaemia mutations were codon 39 and IVS1 nt.110 with the proportions(26.76%) and (20.34%), respectively. No IVS2nt.745 was detected within the studied samples.Genotypic distribution of the samples indicated that there is no significant difference (p > 0.05)between the frequency of homozygotes and heterozygotes within patient group, while there is asignificant difference at (P < 0.01) in comparison with the control group.The study of association between the number and the types of mutations revealed that 28(58%) ofpositive cases have single mutation in a homozygous state or heterozygous state whichsignificantly associated at (P < 0.05) with β-thalassaemia mutations, whereas 20(42%) of thesecases have compound mutations. The most frequent association appeared between IVS1 nt.110 andCodon 39 mutations.Finally, mutations within families, pointed to a positive correlation between the types of mutationsin sons or daughters and their fathers and/or mothers; this indicates the accuracy of the ARMStechnique in detection of β-thalassaemia mutations. This conclusion should be taken with cautiondue to the limited number of families.

Keywords

Beta-thalassaemia --- mutation --- ARMS-PCR --- Iraq


Article
HLA- DRB GENOTYPING OF BRAIN ASTROCYTOMAS AMONG IRAQI PATIENTS
التنميط الجينى لمستضدات خلايا الدم البيضاء الصنف الثانى (HLA-DRB) لسرطان الدماغ النجيمى فى المرضى العراقيين

Authors: Ammira Hadi Al-Shabeeb عامرة هادي --- Nidhal Abdul Mohymen نضال عبد المهيمن محمد
Journal: IRAQI JOURNAL OF MEDICAL SCIENCES المجلة العراقية للعلوم الطبية ISSN: P16816579,E22244719 Year: 2008 Volume: 6 Issue: 2 Pages: 116-120
Publisher: Al-Nahrain University جامعة النهرين

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Background: The major histocompatiblity complex (MHC) refers to as human leukocyte antigen (HLA). The loss of HLA antigens by neoplastic cells is considerably important for tumor growth and metastasis and expression of certain certain HLA allels may predispose to have certain types of tumors. Objective: To investigate the genetic susceptibility of HLA-DRB1, DRB3, DRB4 and. DRB5 alleles to brain astrocytomas in Iraqi patients.Methods: HLA-DRB1, DRB3, DRB4 and, DRB5 allele polymorphisms were typed by polymerase chain- reaction with sequence-specific primers (PCR-SSP) in 30 unrelated patients astrocytomas and 17 unrelated normal control subjects. The association was measured by appropriate statistical tests.Results: Allele frequency (AF) of HLA-DRB1*10011 and DRB1*10012 was significantly decreased in brain astrocytomas patients than that in normal controls (0.53 vs 0.93) the odds ratio 8.76). There was no association between patients and controls in the rested HLA-DRB1 alleles. Conclusion: HLA-DRB1*10011 and DRB1*10012 alleles were less common in the patients with brain astrocytomas than in the healthy controls. Individuals carrying HLA-DRB1* 10011 and DRB1*10012 alleles might be considered as protective markers. These protective alleles; might have a role in the degree of malignancy of the tumors and its histological type.

خلفية الدراسة: تسمى مستضدات التوافق النسيجي (MHC) والخاصة بالخلايا البشرية بـ((HLA.فقدان الخلايا الورميه لهذه المستضدات , تعتبر ظاهرة مهمة لنمو و انتشار الورم . كما إن إنخفاض التعبير عن بعض من اليلات هذه المعقدات يمكن ان يهئ للإصابة بانواع معينة من الأورام.هدف الدراسة: التحري عن امكانية وجود علاقة الإصابة بسرطان الخلايا النجمية في الدماغ وبين وجود اليلات الصنف الثاني من معقدات التوافق النسيجي.المواد و طرائق العمل: تحديد وجود الليلات HLA-DRB1,DRB3,DRB4,DRB5 باستخدام تقنية تفاعل البلمره المتسلسل نوع PCR-SSP)) لدى 30 مريض مصاب بسرطان الخلايا النجمية في الدماغ إضافة إلى 17 شخص سليم كسيطرة.النتائج: وجد ان تكرار اللاليلي لجينات DRB1*10011,DRB1*10012 قد انخفض تواجدهما لدى المرضى مقارنة مع السيطرة (0.5 VS. 0.93). بينما لم نتوصل إلى أي علاقة بين المرض و الاليلات الاخرى.الإستنتاج: من الممكن اعتبار الاليلات DRB1*10011,DRB1*10012 من العوامل التي توفر حمايه ضد الإصابه بهذا الورم.

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