Asking about how can I Isolated DNA from blood parasites like Babesia, Theliria, Plasmodium
Dear Al-Azizz, maybe these papers can help you
http://www.scielo.br/pdf/rimtsp/v55n3/0036-4665-rimtsp-55-03-205.pdf
http://www.scielo.br/pdf/jvatitd/v17n3/03.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4096016/pdf/tru084.pdf
Best regards
Iran J Parasitol. 2016 Oct-Dec;11(4):463-470.
Simplified Pan-species Real-time PCR-based Detection of Plasmodium Spp. in Blood Smear.
Hassanpour G1, Mirhendi H2, Mohebali M1, Raeisi A3, Zeraati H4, Keshavarz H1.
Author information
Abstract
BACKGROUND:
We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan-Plasmodium real-time PCR for accurate screening of individuals suspected of malaria.
METHODS:
A single primer/probe set for pan-species Plasmodium-specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried bloodsmears. FTA cards were used for rapid and simple DNA extraction.
RESULTS:
The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum. All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR.
CONCLUSION:
By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples.
KEYWORDS:
Diagnosis; FTA cards; Malaria; Plasmodium; Real-time PCR
PMID:
28127357
PMCID:
PMC5251174
PLoS Negl Trop Dis. 2016 Dec 1;10(12):e0005146. doi: 10.1371/journal.pntd.0005146. eCollection 2016.
Development of a PCR Assay to Detect Low Level Trypanosoma cruzi in Blood Specimens Collected with PAXgene Blood DNA Tubes for Clinical Trials Treating Chagas Disease.
Wei B1, Chen L1, Kibukawa M1, Kang J2, Waskin H3, Marton M4.
Author information
Abstract
Chagas disease is caused by the parasitic infection of Trypanosoma cruzi (T. cruzi). The STOP CHAGAS clinical trial was initiated in 2011 to evaluate posaconazole in treating Chagas disease, with treatment success defined as negative qualitative PCR results of detecting the parasites in blood specimens collected post-treatment. PAXgene Blood DNA tubes were utilized as a simple procedure to collect and process blood specimens. However, the PAXgene blood specimens challenged published T. cruzi PCR methods, resulting in poor sensitivity and reproducibility. To accurately evaluate the treatment efficacy of the clinical study, we developed and validated a robust PCR assay for detecting low level T. cruzi in PAXgene blood specimens. The assay combines a new DNA extraction method with a custom designed qPCR assay, resulting in limit of detection of 0.005 and 0.01 fg/μl for K98 and CL Brener, two representative strains of two of T. cruzi's discrete typing units. Reliable qPCR standard curves were established for both strains to measure parasite loads, with amplification efficiency ≥ 90% and the lower limit of linearity ≥ 0.05 fg/μl. The assay successfully analyzed the samples collected from the STOP CHAGAS study and may prove useful for future global clinical trials evaluating new therapies for asymptomatic chronic Chagas disease.
PMID:
27906977
PMCID:
PMC5131911
DOI:
10.1371/journal.pntd.0005146
Dear Suzan Al-Azizz
The protocol of Isolation of DNA from blood parasites is easy and you can get much DNA by using High Pure PCR Template Preparation Kit . Briefly, add 200 uL of whole blood on EDTA (fresh or not more than 3 days in refrigerator), then add 200 uL binding buffer and 40 uL Proteinase K, mix immediately, incubate for 10 min at +70°C. Add 100 µl Isopropanol and mix well and completing according to the protocol, which can download from https://lifescience.roche.com/documents/High-Pure-PCR-Template-Preparation-Kit.pdf.
This method is well tested and of high DNA yielded.
Kind regards
The improved DNA extraction kit (QIAamp DNA Mini Kit) was used to extract DNA from blood smears of blood parasites as plasmodium,
Hi ...
Isolation of DNA of blood parasites is easy and you can get much DNA by using extraction Kit
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