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Register a new userTesting 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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Damascus University ورقة بحثية
Publication date
2012
fields
Pharmacy
and research's language is
العربية
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Shamra Editor
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تحتل الإصابات الفطرية المسببة بأنواع فطر الرشاشية Aspergillus عمومـاً، المرتبـة الثانيـة عالمياً بين الإصابات الفطرية الغازيـة، و بـشكل خـاص، تعـد الرشاشـية الـدخناء Aspergillus fumigatus المسبب الرئيسي لداء الرشاشيات الغازي (IA (Aspergillosis Invasive . و على الرغم من أن هذا الداء نادراً ما يصيب الأفراد سليمي المناعة، إلا أنه يؤدي إلى الوفاة لـدى الأشخاص المضعفين مناعياً، لذلك فمن الضروري تشخيصه بشكلٍ مبكر، لمعالجتـه بـشكلٍ نـاجع. إن الأدوات التشخيصية التقليدية المتبعة حالياً للكشف عن الإصابات هي الاستنبات على وسط سـابورو، إذ تتصف هذه الطريقة بأنها ذات حساسية منخفضة و موثوقية ضعيفة. و قد سمح لنا انتشار تقانة التفاعل السلـسلي للبـوليميراز PCR-Nested كتقانـة جزيئيـة ذات حساسية عالية، بإجراء دراسة مقارنة بين حساسية كلتا الطريقتين التقليدية المتبعة و PCR-Nested، 6 و أظهرت نتائج الدراسة على عينات قشع مخموجة تجريبياً بتراكيز مختلفـة راوحـت بـين 10 -10 بوغة/مل، حساسية تقانة PCR-Nested و نوعيتها في الكشف عن أدنى التراكيز، مقارنـة بطريقـة التشخيص التقليدية المتمثلة بالاستنبات الفطري على وسط سابورو التي كانت سلبية للتراكيز جميعها.
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.
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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