scholarly journals Comparison of commercial DNA extraction kits for the detection of bacterial genomic DNA from whole-blood samples using a broad-range PCR

Critical Care ◽  
2009 ◽  
Vol 13 (Suppl 4) ◽  
pp. P13
Author(s):  
B Krulova ◽  
E Nemcova ◽  
B Zaloudikova ◽  
P Nemec ◽  
T Freiberger
Keyword(s):  
Dna Extraction ◽  
Whole Blood ◽  
Genomic Dna ◽  
Blood Samples ◽  
Whole Blood Samples

scholarly journals Deteksi Brucella abortus dari Sampel Darah-Utuh dengan Uji Polymerase Chain Reaction Tanpa Ekstraksi DNA

2020 ◽  
Vol 38 (3) ◽  
pp. 222
Author(s):  
David Ardiyanto ◽  
Hastari Wuryastuty ◽  
Raden Wasito
Keyword(s):  
Beef Cattle ◽  
Dna Extraction ◽  
Brucella Abortus ◽  
Whole Blood ◽  
Economic Losses ◽  
Direct Pcr ◽  
Blood Samples ◽  
Pcr Inhibitor ◽  
Whole Blood Samples ◽  
Species Specific

Abstract              Brucellosis is a zoonotic disease that cause a significant economic losses for cattle industries worldwide. A rapid, precise and accurate diagnosis technique for diagnosis of brucellosis in all stages of the infection is definitely required.  Blood-samples are widely used for PCR-based DNA analysis because they are easily collected, handled, and processed. Direct PCR analysis without DNA extraction has been attempted to reduce time and  costs for routine analysis. This approach is promising but is still limited by the presence of PCR inhibitors that is naturally found  in the blood samples. The objective of this study was to compare the effectivity of direct PCR technique with or without DNA extraction for detection of Brucella abortus in the blood samples. Three whole-blood samples from brucella infected dairy cattle and five whole-blood samples  from beef cattle that having abortion were used as samples in this study. A pair of  bcsp31 primers and IS711 primers were used for amplification of genus-specific and species-specific of Brucella.  The results showed that amplicon in the position of 223 bp and 498 bp that are specific for B. abortus were detected from all of the samples that were analyzed on 1.5% agarose gels. Based on the result it could be concluded that direct PCR analyses without DNA extraction is a sensitive, specific, simple, rapid  and inexpensive assay for detecting B. abortus in the whole blood samples for either dairy or beef cattle and therefore it could  improve the existing surveillance and control programs for brucellosis. Keywords : brucellosis; direct PCR; PCR inhibitor; whole-blood sample; without DNA extraction                           Abstrak              Brucellosis adalah penyakit zoonosis yang menyebabkan kerugian ekonomi yang signifikan bagi industri ternak di seluruh dunia. Teknik diagnosis yang cepat, tepat dan akurat yang dapat digunakan untuk diagnosis brucellosis pada semua tahap infeksi sangat diperlukan. Sampel darah banyak digunakan untuk analisis PCR berbasis DNA karena mudah untuk dikoleksi, ditangani, dan diproses. Metoda PCR langsung tanpa didahului dengan ekstraksi DNA dikembangkan dengan tujuan penghematan waktu dan beaya untuk analisa secara rutin. Tehnik ini sangat menjanjikan tetapi memiliki keterbatasan karena adanya senyawa penghambat PCR yang secara alami terkandung di dalam sampel darah . Tujuan dari penelitian ini adalah membandingkan efektifitas antara uji PCR secara langsung dengan ekstraksi dan tanpa ekstraksi DNA untuk deteksi Brucella abortus di dalam darah. Tiga ( 3 ) sampel darah-EDTA yang berasal dari  sapi penderita brucellosis dan 5 sampel darah-EDTA dari sapi potong yang mengalami abortus digunakan sebagai sampel dalam penelitian ini. Pasangan primer bcsp31 dan primer IS711 untuk amplifikasi gen dan species specific digunakan dalam penelitian. Hasil menunjukkan bahwa amplikon/pita pada posisi 223 bp dan 498 bp yang spesifik untuk Brucella abortus terdeteksi dari semua sampel yang dianalisa dengan gel agarosa 1,5%. Berdasarkan hasil penelitian dapat disimpulkan bahwa uji PCR secara langsung tanpa didahului dengan ekstraksi DNA merupakan tehnik yang sensitif, spesifik, sederhana, cepat dan murah untuk deteksi B. abortus di dalam sampel darah baik sapi perah maupun sapi potong dan oleh karena itu diharapkan dapat digunakan untuk memperbaiki program kontrol dan survailance yang telah ada untuk brucellosis. Kata kunci : brucellosis; PCR langsung; penghambat PCR; sampel darah-utuh; tanpa ekstraksi DNA

scholarly journals An efficient and novel technology for the extraction of parasite genomic DNA from whole blood or culture

BioTechniques ◽  
2020 ◽  
Vol 68 (2) ◽  
pp. 79-84 ◽  
Author(s):  
David J Clark ◽  
Catherine M Moore ◽  
Marc Flanagan ◽  
Katrien Van Bocxlaer ◽  
Evangelia-Theophano Piperaki ◽  
...  
Keyword(s):  
Whole Blood ◽  
Genomic Dna ◽  
Leishmania Donovani ◽  
Small Volume ◽  
Small Sample ◽  
Blood Samples ◽  
Novel Technology ◽  
Spin Column ◽  
Whole Blood Samples ◽  
Pathogen Dna

The aim of this study was to assess pathogen DNA extraction with a new spin column-based method (DNA-XT). DNA from either whole-blood samples spiked with Plasmodium falciparum or  Leishmania donovani amastigote culture was extracted with DNA-XT and compared with that produced by a commercial extraction kit (DNeasy®). Eluates from large and small sample volumes were assessed by PCR and spectroscopy. Using a small volume (5 μl) of blood, the DNA-XT and DNeasy methods produced eluates with similar DNA concentrations (0.63 vs 1.06 ng/μl, respectively). The DNA-XT method produced DNA with lower PCR inhibition than DNeasy. The new technique was also twice as fast and required fewer plastics and manipulations but had reduced total recovered DNA compared with DNeasy.

Steadiness/stability of antiepileptic drugs in serum, plasma and whole-blood samples under various storage methods

2010 ◽  
Vol 41 (02) ◽  
Author(s):  
N Shazi ◽  
A Böss ◽  
HJ Merkel ◽  
F Scharbert ◽  
D Hannak ◽  
...  
Keyword(s):  
Antiepileptic Drugs ◽  
Whole Blood ◽  
Blood Samples ◽  
Storage Methods ◽  
Whole Blood Samples
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