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The optimization of molecular detection of clinical isolates of brucella in blood cultures by eryD transcriptase gene for confirmation of culture-negative samples
المؤلفون المشاركون
Tabibnejad, Mahsa
Arjomandzadegan, Muhammad
Hashimi, Sayyid Hamid
Nasiri, Zahrah
Alikhani, Muhammad Yusuf
المصدر
Iranian Red Crescent Medical Journal
العدد
المجلد 18، العدد 4 (30 إبريل/نيسان 2016)، ص ص. 1-6، 6ص.
الناشر
تاريخ النشر
2016-04-30
دولة النشر
الإمارات العربية المتحدة
عدد الصفحات
6
التخصصات الرئيسية
الملخص EN
Background: Brucellosis is a zoonosis disease which is widespread across the world.
Objectives: The aim of the present study is the evaluation of culture-negative blood samples.
Materials and Methods: A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests.
Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method.
Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar.
At thesame time,DNAfrom all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit).
A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions.
The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR.
DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared.
Results: 39 cases (39%) had positive resultswhentested by the BACTEC system, and 61 cases (61%) became negative.
23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene.
Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR.
The specificity of the PCR method was equal to 100%.
DNAextraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples.
Conclusions: The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples.
The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction.
نمط استشهاد جمعية علماء النفس الأمريكية (APA)
Tabibnejad, Mahsa& Alikhani, Muhammad Yusuf& Arjomandzadegan, Muhammad& Hashimi, Sayyid Hamid& Nasiri, Zahrah. 2016. The optimization of molecular detection of clinical isolates of brucella in blood cultures by eryD transcriptase gene for confirmation of culture-negative samples. Iranian Red Crescent Medical Journal،Vol. 18, no. 4, pp.1-6.
https://search.emarefa.net/detail/BIM-682378
نمط استشهاد الجمعية الأمريكية للغات الحديثة (MLA)
Tabibnejad, Mahsa…[et al.]. The optimization of molecular detection of clinical isolates of brucella in blood cultures by eryD transcriptase gene for confirmation of culture-negative samples. Iranian Red Crescent Medical Journal Vol. 18, no. 4 (Apr. 2016), pp.1-6.
https://search.emarefa.net/detail/BIM-682378
نمط استشهاد الجمعية الطبية الأمريكية (AMA)
Tabibnejad, Mahsa& Alikhani, Muhammad Yusuf& Arjomandzadegan, Muhammad& Hashimi, Sayyid Hamid& Nasiri, Zahrah. The optimization of molecular detection of clinical isolates of brucella in blood cultures by eryD transcriptase gene for confirmation of culture-negative samples. Iranian Red Crescent Medical Journal. 2016. Vol. 18, no. 4, pp.1-6.
https://search.emarefa.net/detail/BIM-682378
نوع البيانات
مقالات
لغة النص
الإنجليزية
الملاحظات
Includes bibliographical references : p. 5-6
رقم السجل
BIM-682378
قاعدة معامل التأثير والاستشهادات المرجعية العربي "ارسيف Arcif"
أضخم قاعدة بيانات عربية للاستشهادات المرجعية للمجلات العلمية المحكمة الصادرة في العالم العربي
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