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Article
Epidemiological, Clinical, and immunological characteristics of Mycoplasma pneumonia infections among a group of hospitalized children in Suleimani city/Iraq .

Author: Ali H. bayati*, Pakhshan M. Faraj*, Khalid H. Salih**
Journal: Al-Kindy College Medical Journal مجلة كلية الطب الكندي ISSN: 18109543 Year: 2014 Volume: 10 Issue: 2 Pages: 5-9
Publisher: Baghdad University جامعة بغداد

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Abstract

Background: Mycoplasma pneumoniae (M. pneumoniae) is an important respiratory bacterial pathogen, especially among children. It causes acute upper and lower respiratory infections.Objective: This study was aimed to measure anti- M. pneumoniae antibodies among hospitalized children who were admitted to hospital diagnosed with acute respiratory tract infections.Method: Automated ELISA technique was performed to detect anti- M. pneumoniae antibodies (IgM and IgG antibodies) in serum from 108 children less than 5 years old. The children were admitted to the Pediatric Teaching Hospital in Suleimani city/Kurdistan Region/Iraq because of acute respiratory tract infections. A questionnaire was designed to collect demographic and clinical data from those children.Results: IgM anti- M. pneumoniae antibodies were positive in 15 (13.9%) out of 108 children. The highest seroprevalence was found in the age group 25-36 months while the lowest is in the age group 1-12 months. M. pneumoniae infections were more common among males than females though results were statistically not significant, and attendance of kindergarten or nursery, residency, history of chronic diseases, history of contact with similar conditions, and family history of chronic diseases, they were all statistically not significant. The IgMseropositive children were suffering from bronchitis, croup, pneumonia, or other respiratory infections, in frequencies of 7 (46.6%), 4 (26.7%), 3 (20%), and 1 (6.7%) respectively. Increased erythrocyte sedimentation rate, diagnosis of croup, and diagnosis of bronchitis were more frequent in M. pneumoniae infected group and the results were statistically significant. The IgG anti- M. pneumoniae antibodies were positive in 31 (28.8%) out of the 108 children, and the greatest IgG seroprevalence was highest in age group 49-60 months.Conclusion: M. pneumoniae is an important respiratorypathogen among hospitalized children in Sulaimanigovernorate/Kurdistan/Iraq, and nearly one third of childrenhad experienced M. pneumoniae infection by the age of fiveyears


Article
Advanced and conventional Molecular techniques in the diagnosis of Mycoplasma pneumoniae in patients with pneumoniae
تقنيات بيولوجية جزيئية متقدمة و تقليدية للكشف عن بكتريا المفطورات الرئوية في المرضى المصابين بالتهاب القناة التنفسية

Authors: Nidhal A.Mohammed نضال عبد المهيمن --- Mushtak T.S. Al-Ouqaili مشتاق طالب صالح --- Muntaha M.Hassan منتهى مداح حسن
Journal: Al- Anbar Medical Journal مجلة الأنبار الطبية ISSN: PISSN: 27066207 / EISSN: 26643154 Year: 2013 Volume: 11 Issue: 1 Pages: 37-46
Publisher: University of Anbar جامعة الانبار

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Abstract

Background:-M. pneumoniae is an important human pathogen that produces community-acquired respiratory tract infection. Diagnosis of M. pneumoniae infection is challenging and crucial for the timely initiation of the effective antibiotic therapy.Objective: This study has been undertaken to detect M. pneumoniae in respiratory samples (throat swabs, throat wash and sputum) in patients with respiratory tract infection qualitatively by conventional polymerase chain reaction (PCR). Also, more advanced one, real time PCR was used to determine mycoplasmal target gene qualitatively and quantitatively. Patients and methods: The study was performed on Seventy-five patients and thirty healthy subjects as control. Human genomic DNA was extracted and M. pneumoniae target gene (lipoprotein gene) was amplified using conventional PCR. Negative, positive controls and internal controls were involved in each experimental run. The amplified products were analyzed in 2% agarose gel and visualized using Red safe staining. In real time PCR, specific primer and probe mix depending on TaqMan® principle was used to detect P1 adhesion gene through FAM channel. A fluorogenic probe was included in the same reaction mixture which consists of a DNA probe labeled with a 5`-dye and a 3`-quencher. During PCR amplification. Data were analyzed using Smart-cycler software and M. pneumoniae DNA copy number was estimated from the cross point threshold relative to positive standard.Results: Thirty five patients (45.5%) were positive by PCR and Thirty two (42.6 %) were positive by Real-time PCR. The highest rate of infection by using two molecular methods was of less than 20 years of age. The quantity of M. pneumoniae DNA target gene in positive Real-time PCR was ranged between 10-2000copies/µl.Conclusion: The study concluded that both of molecular techniques conventional and real-time PCR are a rapid, reliable and ideal in diagnosis of M. pneumoniae using throat swabs, throat wash and sputum samples

Background:-M. pneumoniae is an important human pathogen that produces community-acquired respiratory tract infection. Diagnosis of M. pneumoniae infection is challenging and crucial for the timely initiation of the effective antibiotic therapy.Objective: This study has been undertaken to detect M. pneumoniae in respiratory samples (throat swabs, throat wash and sputum) in patients with respiratory tract infection qualitatively by conventional polymerase chain reaction (PCR). Also, more advanced one, real time PCR was used to determine mycoplasmal target gene qualitatively and quantitatively. Patients and methods: The study was performed on Seventy-five patients and thirty healthy subjects as control. Human genomic DNA was extracted and M. pneumoniae target gene (lipoprotein gene) was amplified using conventional PCR. Negative, positive controls and internal controls were involved in each experimental run. The amplified products were analyzed in 2% agarose gel and visualized using Red safe staining. In real time PCR, specific primer and probe mix depending on TaqMan® principle was used to detect P1 adhesion gene through FAM channel. A fluorogenic probe was included in the same reaction mixture which consists of a DNA probe labeled with a 5`-dye and a 3`-quencher. During PCR amplification. Data were analyzed using Smart-cycler software and M. pneumoniae DNA copy number was estimated from the cross point threshold relative to positive standard.Results: Thirty five patients (45.5%) were positive by PCR and Thirty two (42.6 %) were positive by Real-time PCR. The highest rate of infection by using two molecular methods was of less than 20 years of age. The quantity of M. pneumoniae DNA target gene in positive Real-time PCR was ranged between 10-2000copies/µl.Conclusion: The study concluded that both of molecular techniques conventional and real-time PCR are a rapid, reliable and ideal in diagnosis of M. pneumoniae using throat swabs, throat wash and sputum samples


Article
Advanced and conventional molecular techniques in the diagnosis of Mycoplasma pneumoniae in patients with respiratory tract infection
تقنيات بيولوجية جزيئية متقدمة و تقليدية للكشف عن بكتريا المفطورات الرئوية في المرضى المصابين بالتهاب القناة التنفسية

Authors: Muntaha M. Hassan منتهى مداح حسن --- Mushtak T.S. Al-Ouqaili مشتاق طالب صالح --- Nidhal A. Mohammed نضال عبد المهيمن --- Omar A. Mohammed عمر علي محمد
Journal: Journal of the Faculty of Medicine مجلة كلية الطب ISSN: PISSN: 00419419 / EISSN: 24108057 Year: 2014 Volume: 56 Issue: 1 Pages: 95-100
Publisher: Baghdad University جامعة بغداد

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Abstract

Background:-M. pneumoniae is an important human pathogen that produces community-acquired respiratory tract infection. Diagnosis of M. pneumoniae infection is challenging and crucial for the timely initiation of the effective antibiotic therapy.Objective: This study has been undertaken to detect M. pneumoniae in respiratory samples (throat swabs, throat wash and sputum) in patients with respiratory tract infection qualitatively by conventional polymerase chain reaction (PCR). Also, more advanced one, real time PCR was used to determine mycoplasmal target gene qualitatively and quantitatively. Patients and methods: The study was performed on Seventy-five patients and thirty healthy subject as control. Genomic DNA was extracted and M. pneumoniae target gene (lipoprotein gene) was amplified using conventional PCR. Negative, positive controls and internal controls were involved in each experimental run. The amplified products were analyzed in 2% agarose gel and visualized using Red safe staining. In real time PCR, specific primer and probe mix depending on TaqMan® principle was used to detect P1 adhesion gene through FAM channel. A fluorogenic probe was included in the same reaction mixture which consists of a DNA probe labeled with a 5`-dye and a 3`-quencher. During PCR amplification Data were analyzed using Smart-cycler software and M. pneumoniae DNA copy number was estimated from the cross point threshold relative to positive standard.Results: Thirty five patients(45.5%) were positive by PCR and Thirty two (42.6 %) were positive by Real-time PCR. The highest rate of infection by using tow molecular methods were of less than 20 years of age. The quantity of M. pneumoniae DNA target gene in positive Real-time PCR were ranged between 10-2000copies/µl.Conclusion: The study concluded that both of molecular techniques conventional and real-time PCR are a rapid, reliable and ideal in diagnosis of M. pneumoniae using throat swabs, throat wash and sputum samples.Key word: Mycoplasma pneumoniae, PCR , Real-time PCR

الخلاصة:خلفية الدراسة :- بكتريا المفطورات الرئوية من الممرضات المهمة للإنسان والتي تسبب إصابات القناة التنفسية المكتسبة من المجتمع. تشخيص الإصابة بهذه البكتريا ليست سهلة وكذلك حاسمة من اجل اختيار الوقت المناسب للبدء بالمضادات الحيوية.هدف الدراسة:- صممت هذه الدراسة للكشف عن بكتريا المفطورات الرئوية في العينات المأخوذة من القناة التنفسية (مسحة الحنجرة, غسيل الحنجرة, البلغم) للمرضى المصابين بالتهاب القناة التنفسية باستخدام الفحوصات النوعية كتفاعل البلمرة المتسلسل التقليدي. كذلك للفحص الكمي لتفاعل البلمرة المتسلسل للكشف عن وتقييس ألجين الهدف لبكتريا المفطورات الرئوية نوعيا وكميا.المرضى وطرق العمل:-أنجزت الدراسة على 75مريضا و30 شخصا سليما كعينة للسيطرة. تم استخلاص الحامض النووي ( دنا) من العينات التنفسية البشرية , والجين (lipoprotein gene ) تم تضخيمه باستخدام تفاعل البلمرة المتسلسل التقليدي. فحص السيطرة تم إدراجه في كل تجربة. الناتج المضخم تم الكشف عنه بالترحيل الكهربائي باستخدام جل الاكاروز (2% ) وتم توضيحه باستخدام صبغة ( Red safe ). تم استخدام برايمر وبروب خاص بتقنية ( TaqMan®) من خلال قناة. FAM البيانات تم تحليلها باستخدام برامج للعقل الإلكتروني الخاصة بجهاز Smart-cycler, عدد نسخ الحامض النووي منقوص الأوكسجين دنا تم تقديرها من خلال نقاط التقاطع بين حد العتبة لكل دورة و التراكيز القياسية.النتائج: أربعة وثلاثين مريضا (45.5% ) أظهروا نتائج موجبة لفحص البلمرة المتسلسل, بينما اظهر 32 مريضا ( 42.6 %) نتيجة موجبة للفحص الكمي لتفاعل البلمرة المتسلسل. النسبة الأعلى للإصابة باستخدام كلا الفحصين الجزيئيين كانت للفئة العمرية الأقل من 20 سنة. كمية الحامض النووي دنا لبكتريا المفطورات الرئوية في النماذج التي أعطت نتيجة موجبة للفحص الكمي لتفاعل البلمرة المتسلسل تراوحت بين 10-2000 نسخة / مايكرولتر.الاستنتاجات:- تبين من الدراسة الحالية ان كلا من الفحوص الجزيئية المستخدمة كتفاعل البلمرة المتسلسل التقليدي و الفحص الكمي لتفاعل البلمرة المتسلسل تمتاز بأنها سريعة, موثوقة وكذلك مناسبة لتشخيص بكتريا المفطورات الرئوية باستخدام العينات التنفسية (مسحة الحنجرة, غسيل الحنجرة, البلغم).مفتاح الكلمات: بكتريا المفطورات الرئوية, فحص البلمرة المتسلسل, الفحص الكمي لتفاعل البلمرة المتسلسل

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