uu.seUppsala University Publications
Change search
ReferencesLink to record
Permanent link

Direct link
SYBR Green Real-Time PCR-RFLP Assay Targeting the Plasmodium Cytochrome B Gene - A Highly Sensitive Molecular Tool for Malaria Parasite Detection and Species Determination
Show others and affiliations
2015 (English)In: PLoS ONE, ISSN 1932-6203, Vol. 10, no 3, e0120210Article in journal (Refereed) Published
Abstract [en]

A prerequisite for reliable detection of low-density Plasmodium infections in malaria pre-elimination settings is the availability of ultra-sensitive and high-throughput molecular tools. We developed a SYBR Green real-time PCR restriction fragment length polymorphism assay (cytb-qPCR) targeting the cytochrome b gene of the four major human Plasmodium species (P. falciparum, P. vivax, P. malariae, and P. ovale) for parasite detection and species determination with DNA extracted from dried blood spots collected on filter paper. The performance of cytb-qPCR was first compared against four reference PCR methods using serially diluted Plasmodium samples. The detection limit of the cytb-qPCR was 1 parasite/mu l (p/mu l) for P. falciparum and P. ovale, and 2 p/mu l for P. vivax and P. malariae, while the reference PCRs had detection limits of 0.5-10 p/mu l. The ability of the PCR methods to detect low-density Plasmodium infections was then assessed using 2977 filter paper samples collected during a cross-sectional survey in Zanzibar, a malaria pre-elimination setting in sub-Saharan Africa. Field samples were defined as 'final positive' if positive in at least two of the five PCR methods. Cytb-qPCR preformed equal to or better than the reference PCRs with a sensitivity of 100% (65/65; 95% CI 94.5-100%) and a specificity of 99.9%(2910/2912; 95% CI 99.7-100%) when compared against 'final positive' samples. The results indicate that the cytb-qPCR may represent an opportunity for improved molecular surveillance of low-density Plasmodium infections in malaria pre-elimination settings.

Place, publisher, year, edition, pages
2015. Vol. 10, no 3, e0120210
National Category
Infectious Medicine Cell Biology
URN: urn:nbn:se:uu:diva-252009DOI: 10.1371/journal.pone.0120210ISI: 000351183500158PubMedID: 25774805OAI: oai:DiVA.org:uu-252009DiVA: diva2:810447
Available from: 2015-05-07 Created: 2015-04-28 Last updated: 2015-05-07Bibliographically approved

Open Access in DiVA

fulltext(3141 kB)67 downloads
File information
File name FULLTEXT01.pdfFile size 3141 kBChecksum SHA-512
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed
By organisation
Centrum för klinisk forskning i Sörmland (CKFD)
In the same journal
Infectious MedicineCell Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 67 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 216 hits
ReferencesLink to record
Permanent link

Direct link