استفاده از Nested-PCR در مطالعه انتشار انگل Cryptosporidium spp در پرندگان اهلی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 دانشکده پرستاری، دانشگاه القادسیه، عراق

2 دانشکده بیوتکنولوژی، دانشگاه القادسیه، عراق

3 دانشکده علوم، دانشگاه القادسیه، عراق

چکیده

هدف: ین مطالعه به بررسی تعداد موارد عفونت Cryptosporidium spp. در پرندگان اهلی در استان دیوانیه و همچنین پیامدهای این بیماری و روش های تشخیص آن پرداخت.
مواد و روش‌ها: در این مطالعه، 205 نمونه از گونه های مختلف پرنده بدون در نظر گرفتن جنسیت مورد تجزیه و تحلیل قرار گرفت. برخی از روش‌های مورد استفاده برای تشخیص، روش رنگ آمیزی اسید فست و تکنیک نستد PCR بود. ثابت شد که نستد PCR بهتر از رنگ آمیزی اسید فست AFS است. زیرا می‌تواند کریپتوسپوریدیوم را سریعتر، با بالاترین دقت و حساسیت شناسایی کند.
نتایج: نتایج نشان داد که 90 نمونه از کل نمونه‌ها دارای عفونت کریپتوسپوریدیوم بودند. در بین گونه های پرنده مورد مطالعه، falseco eleonorae  با 16% کمترین بروز کریپتوسپوریدیوزیس را داشت، در حالی که گالوس گالوس داخلی با 3/53 درصد بیشترین بروز را داشت. از 60 نمونه گرفته شده برای هر گونه، 3/48 درصد از گونه Meleagris gallopova و 6/41 درصد از گونه Anas platyrhynchos آلوده بودند. بررسی میکروسکوپی میزان عفونت 9/43 درصد (90 نمونه از 205 نمونه) را در مقایسه با روش دیگر نشان داد. برای استفاده از Nested-PCR برای شناسایی مولکولی، DNA باید از نمونه های آلوده مثبت با استفاده از روش پروتئیناز K استخراج شود. این کار باندهای مثبتی را تولید کرد که معمولاً در محدوده 587 جفت بازی تکثیر می‌شدند. تست Nested-PCR نشان داد که 5/47 درصد نمونه ها (19 از 40 نمونه) آلوده بودند.
نتیجه‌گیری: در نهایت، این مطالعه با نشان دادن شیوع بسیار بالای عفونت کریپتوسپوریدیوم در پرندگان اهلی در استان دیوانیه، نیاز حیاتی به ابزارهای تشخیصی کارآمد و طرح‌های مداخله‌ای را برجسته کرد. به منظور جلوگیری از گسترش کریپتوسپوریدیوزیس در پرندگان، نتایج بر استفاده از روش‌های واکنش زنجیره‌ای پلیمراز نستد (nested-PCR) برای تشخیص عفونت تاکید دارد.

کلیدواژه‌ها


عنوان مقاله [English]

Using Nested-PCR in the Study of Cryptosporidium spp Parasite Spread in Domestic Bird

نویسندگان [English]

  • Saafa Ressan Abdullah Al-Kaeebi 1
  • Rana Saleh Al-Difaie 2
  • Fatima Ibrahim Mohammad 3
1 College of Nursing College, University of Al-Qadisiyah, Iraq
2 Biotechnology College, University of Al-Qadisiyah, Iraq.
3 College of science, University of Al-Qadisiyah, Iraq
چکیده [English]

Objectives
This study examined the number of cases of Cryptosporidium spp. infection in domestic birds in Diwaniyah Governorate, as well as the consequences of this illness and the methods used to diagnose it.
 
Materials and Methods
In the study, 205 samples from different bird species were analyzed without gender being taken into account. Some of the methods used for diagnosis were the Acid Fast Stain method and the DNA nested Polymerization Technique (PCR). Nested - PCR proved out to be better than AFS because it could find Cryptosporidium faster, with highest accuracy, and sensitivity.
 
Results
The results showed that 90 of the samples had Cryptosporidium infections. Among the bird species that were studied, falseco eleonorae had the lowest incidence of cryptosporidiosis at 16%, while gallus gallus domesticus had the highest incidence at 53.3%. Out of the 60 samples taken for each species, 48.3% of Meleagris gallopova and 41.6% of Anas platyrhynchos were infected. The microscope examination showed a 43.9% infection rate (90 out of 205 samples) when compared to the other method. To use nested-PCR for molecular identification, DNA had to be taken from positive infected samples using Proteinase K method. This produced positive bands that were usually amplified at around 587 bp. The nested-PCR test showed that 47.5% of the samples (19 out of 40) were infected.
 
Conclusions
Finally, this study highlights the critical need for efficient diagnostic tools and intervention plans by demonstrating the very high incidence of Cryptosporidium infection in domestic birds in Diwaniyah Governorate. In order to prevent the spread of Cryptosporidiosis in birds, the results stress the use of nested polymerase chain reaction (PCR) methods for infection diagnosis.

کلیدواژه‌ها [English]

  • Nested-PCR
  • Microscopic Examination
  • Infection Rate
  • Birds
  • and Cryptosporidium
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