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Testing the sensitivity of Nested PCR method to detect Aspergillus fumigates in experimentally infected Sputum samples

دراسة حساسية تقانة PCR Nested في كشف الرشاشية الدخناء في عينات قشع مخموجة تجريبي

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 Publication date 2012
  fields Pharmacy
and research's language is العربية
 Created by Shamra Editor




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Fungal infections caused by Aspergillus species generally are occupying a second place among invasive fungal infections in the world, especially A. fumigatus, which is considered the main cause of invasive Aspergillosis (IA). Although IA rarely infects immunocompetent individuals, however, it can lead to death in immunocompromised patients. Therefore, it is necessary to diagnose the infection early in order to treat the disease efficiently. However, the conventional diagnostic tools, currently used to detect infections, has low sensitivity and reliability. Polymerase chain reaction (PCR) technology distribution as a molecular and high sensitive technology has allowed us to make comparative study between sensitivity of traditional currently used diagnostic method and Nested-PCR, the result of the study of sputum samples that experimentally infected with different concentrations of A.fumigatus spores ramping from 10 to106 spore/ml, have high sensitivity and specificity of Nested-PCR in detecting the lower concentrations, comparing with traditional diagnostic method (culture on Sabouraud media) that were negative in all concentrations.


Artificial intelligence review:
Research summary
تتناول الدراسة حساسية تقانة Nested PCR في كشف فطر الرشاشية الدخناء في عينات القشع المخموجة تجريبياً. تعتبر الإصابات الفطرية المسببة بفطر الرشاشية من الإصابات الخطيرة التي تؤدي إلى الوفاة لدى الأشخاص المضعفين مناعياً. تعتمد الطرق التقليدية للكشف عن هذه الإصابات على الاستنبات الفطري على وسط سابورو، والتي تتميز بحساسية منخفضة وموثوقية ضعيفة. في هذه الدراسة، تم مقارنة حساسية تقانة Nested PCR مع الطريقة التقليدية للكشف عن فطر الرشاشية الدخناء. أظهرت النتائج أن تقانة Nested PCR تتميز بحساسية ونوعية عالية في الكشف عن الفطر حتى في التراكيز المنخفضة، بينما كانت نتائج الاستنبات الفطري سلبية لجميع التراكيز المستخدمة. تم الحصول على عينات القشع من شخص سليم، وتم تخميجها بتراكيز مختلفة من أبواغ الفطر. تم استنبات العينات على وسط سابورو كلورامفينيكول، واستخلص DNA الجينومي الفطري من العينات وأجريت عليها تقانة Nested PCR. أظهرت النتائج أن تقانة Nested PCR قادرة على الكشف عن الفطر في جميع التراكيز المستخدمة، مما يبرز فعاليتها العالية مقارنة بالطريقة التقليدية.
Critical review
دراسة نقدية: تعتبر هذه الدراسة خطوة مهمة نحو تحسين طرق التشخيص للإصابات الفطرية الغازية، وخاصة داء الرشاشيات. ومع ذلك، يمكن توجيه بعض النقد البناء للدراسة. أولاً، كان من الممكن توسيع حجم العينة لتشمل مزيداً من العينات من مرضى حقيقيين بدلاً من العينات المخموجة تجريبياً، مما يعزز من موثوقية النتائج وتطبيقها في الواقع العملي. ثانياً، لم تتناول الدراسة بشكل كافٍ العوامل المحتملة التي قد تؤثر على حساسية تقانة Nested PCR، مثل وجود ملوثات أو تداخلات من مكونات أخرى في العينات. أخيراً، كان من الممكن مقارنة تقانة Nested PCR مع تقانات جزيئية أخرى مثل Real-Time PCR لتقديم صورة أشمل عن أفضل الطرق المتاحة للكشف عن فطر الرشاشية الدخناء.
Questions related to the research
  1. ما هي الفطريات التي تسبب داء الرشاشيات الغازي؟

    الفطريات التي تسبب داء الرشاشيات الغازي هي أنواع فطر الرشاشية، وخاصة الرشاشية الدخناء (Aspergillus fumigatus).

  2. ما هي الطريقة التقليدية المستخدمة للكشف عن فطر الرشاشية الدخناء؟

    الطريقة التقليدية المستخدمة للكشف عن فطر الرشاشية الدخناء هي الاستنبات الفطري على وسط سابورو كلورامفينيكول.

  3. ما هي مزايا تقانة Nested PCR مقارنة بالطريقة التقليدية؟

    تتميز تقانة Nested PCR بحساسية ونوعية عالية في الكشف عن فطر الرشاشية الدخناء حتى في التراكيز المنخفضة، مقارنة بالطريقة التقليدية التي تتميز بحساسية منخفضة وموثوقية ضعيفة.

  4. ما هي العوامل التي قد تؤثر على حساسية تقانة Nested PCR؟

    العوامل التي قد تؤثر على حساسية تقانة Nested PCR تشمل وجود ملوثات أو تداخلات من مكونات أخرى في العينات، وكذلك جودة ونقاء DNA المستخلص.


References used
Araujo R. & Rodrigues A. G. (2004). Variability Of Germinative Potential among Pathogenic Species of Aspergillus. J. Clin Microbiol 42, 4335-4337
Bansod S., Gupta I. & Rai M. (2008). Specific detection of Aspergillus fumigatus in sputum sample of pulmonary tuberculosis patients by twostep PCR. African Journal of Biotechnology 7, 16-21
Bodey G., Bueltmann B. & Duguid W., et al.. (1992). Fungal infections in cancer patients: an international autopsy survey. Eur. J. Clin Microbiol Infect Dis 11, 99-109
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