PubTransformer

A site to transform Pubmed publications into these bibliographic reference formats: ADS, BibTeX, EndNote, ISI used by the Web of Knowledge, RIS, MEDLINE, Microsoft's Word 2007 XML.

Four human Plasmodium species quantification using droplet digital PCR.

Abstract Droplet digital polymerase chain reaction (ddPCR) is a partial PCR based on water-oil emulsion droplet technology. It is a highly sensitive method for detecting and delineating minor alleles from complex backgrounds and provides absolute quantification of DNA targets. The ddPCR technology has been applied for detection of many pathogens. Here the sensitive assay utilizing ddPCR for detection and quantification of Plasmodium species was investigated. The assay was developed for two levels of detection, genus specific for all Plasmodium species and for specific Plasmodium species detection. The ddPCR assay was developed based on primers and probes specific to the Plasmodium genus 18S rRNA gene. Using ddPCR for ultra-sensitive P. falciparum assessment, the lower level of detection from concentrated DNA obtained from a high volume (1 mL) blood sample was 11 parasites/mL. For species identification, in particular for samples with mixed infections, a duplex reaction was developed for detection and quantification P. falciparum/ P. vivax and P. malariae/ P. ovale. Amplification of each Plasmodium species in the duplex reaction showed equal sensitivity to singleplex single species detection. The duplex ddPCR assay had higher sensitivity to identify minor species in 32 subpatent parasitaemia samples from Cambodia, and performed better than real-time PCR. The ddPCR assay shows high sensitivity to assess very low parasitaemia of all human Plasmodium species. This provides a useful research tool for studying the role of the asymptomatic parasite reservoir for transmission in regions aiming for malaria elimination.
PMID
Related Publications

Comparison of diagnostic methods for the detection and quantification of the four sympatric Plasmodium species in field samples from Papua New Guinea.

Evaluation of FRET real-time PCR assay for rapid detection and differentiation of Plasmodium species in returning travellers and migrants.

Detection and identification of human Plasmodium species with real-time quantitative nucleic acid sequence-based amplification.

Plasmodium malariae and Plasmodium ovale infections in the China-Myanmar border area.

Towards high-throughput molecular detection of Plasmodium: new approaches and molecular markers.

Authors

Mayor MeshTerms
Keywords
Journal Title plos one
Publication Year Start




PMID- 28423028
OWN - NLM
STAT- In-Process
DA  - 20170419
LR  - 20170419
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 12
IP  - 4
DP  - 2017
TI  - Four human Plasmodium species quantification using droplet digital PCR.
PG  - e0175771
LID - 10.1371/journal.pone.0175771 [doi]
AB  - Droplet digital polymerase chain reaction (ddPCR) is a partial PCR based on
      water-oil emulsion droplet technology. It is a highly sensitive method for
      detecting and delineating minor alleles from complex backgrounds and provides
      absolute quantification of DNA targets. The ddPCR technology has been applied for
      detection of many pathogens. Here the sensitive assay utilizing ddPCR for
      detection and quantification of Plasmodium species was investigated. The assay
      was developed for two levels of detection, genus specific for all Plasmodium
      species and for specific Plasmodium species detection. The ddPCR assay was
      developed based on primers and probes specific to the Plasmodium genus 18S rRNA
      gene. Using ddPCR for ultra-sensitive P. falciparum assessment, the lower level
      of detection from concentrated DNA obtained from a high volume (1 mL) blood
      sample was 11 parasites/mL. For species identification, in particular for samples
      with mixed infections, a duplex reaction was developed for detection and
      quantification P. falciparum/ P. vivax and P. malariae/ P. ovale. Amplification
      of each Plasmodium species in the duplex reaction showed equal sensitivity to
      singleplex single species detection. The duplex ddPCR assay had higher
      sensitivity to identify minor species in 32 subpatent parasitaemia samples from
      Cambodia, and performed better than real-time PCR. The ddPCR assay shows high
      sensitivity to assess very low parasitaemia of all human Plasmodium species. This
      provides a useful research tool for studying the role of the asymptomatic
      parasite reservoir for transmission in regions aiming for malaria elimination.
FAU - Srisutham, Suttipat
AU  - Srisutham S
AD  - Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical
      Medicine, Mahidol University, Bangkok, Thailand.
FAU - Saralamba, Naowarat
AU  - Saralamba N
AD  - Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical
      Medicine, Mahidol University, Bangkok, Thailand.
AD  - Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine,
      Mahidol University, Bangkok, Thailand.
FAU - Malleret, Benoit
AU  - Malleret B
AD  - Laboratory of Pathogen Immunobiology, Singapore Immunology Network (SIgN), Agency
      for Science Technology and Research (A*STAR), Biopolis, Singapore, Singapore.
AD  - Department of Microbiology and Immunology, Yong Loo Lin School of Medicine,
      National University of Singapore, National University Health System, Singapore,
      Singapore.
FAU - Renia, Laurent
AU  - Renia L
AD  - Laboratory of Pathogen Immunobiology, Singapore Immunology Network (SIgN), Agency
      for Science Technology and Research (A*STAR), Biopolis, Singapore, Singapore.
FAU - Dondorp, Arjen M
AU  - Dondorp AM
AD  - Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine,
      Mahidol University, Bangkok, Thailand.
AD  - Centre for Tropical Medicine and Global Health, Churchill Hospital, University of
      Oxford, Oxford, United Kingdom.
FAU - Imwong, Mallika
AU  - Imwong M
AUID- ORCID: http://orcid.org/0000-0003-0857-1855
AD  - Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical
      Medicine, Mahidol University, Bangkok, Thailand.
AD  - Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine,
      Mahidol University, Bangkok, Thailand.
LA  - eng
PT  - Journal Article
DEP - 20170419
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
EDAT- 2017/04/20 06:00
MHDA- 2017/04/20 06:00
CRDT- 2017/04/20 06:00
PHST- 2017/02/01 [received]
PHST- 2017/03/30 [accepted]
AID - 10.1371/journal.pone.0175771 [doi]
AID - PONE-D-17-04245 [pii]
PST - epublish
SO  - PLoS One. 2017 Apr 19;12(4):e0175771. doi: 10.1371/journal.pone.0175771.
      eCollection 2017.

<?xml version="1.0" encoding="UTF-8"?>
<b:Sources SelectedStyle="" xmlns:b="http://schemas.openxmlformats.org/officeDocument/2006/bibliography"  xmlns="http://schemas.openxmlformats.org/officeDocument/2006/bibliography" >
</b:Sources>