PMID- 26743662
OWN - NLM
STAT- MEDLINE
DCOM- 20161226
LR  - 20181202
IS  - 1365-2672 (Electronic)
IS  - 1364-5072 (Linking)
VI  - 120
IP  - 5
DP  - 2016 May
TI  - High-throughput multiplex real-time PCR assay for the simultaneous quantification 
      of DNA and RNA viruses infecting cassava plants.
PG  - 1346-56
LID - 10.1111/jam.13043 [doi]
AB  - AIMS: To develop a multiplex TaqMan-based real-time PCR assay (qPCR) for the 
      simultaneous detection and quantification of both RNA and DNA viruses affecting 
      cassava (Manihot esculenta) in eastern Africa. METHODS AND RESULTS: The 
      diagnostic assay was developed for two RNA viruses; Cassava brown streak virus 
      (CBSV) and Uganda cassava brown streak virus (UCBSV) and two predominant DNA 
      viruses; African cassava mosaic virus (ACMV) and East African cassava mosaic 
      virus (EACMV), which cause the economically important cassava brown streak 
      disease (CBSD) and cassava mosaic disease (CMD) respectively. Our method, 
      developed by analysing PCR products of viruses, was highly sensitive to detect 
      target viruses from very low quantities of 4-10 femtograms. Multiplexing did not 
      diminish sensitivity or accuracy compared to uniplex alternatives. The assay 
      reliably detected and quantified four cassava viruses in field samples where CBSV 
      and UCBSV synergy was observed in majority of mixed-infected varieties. 
      CONCLUSIONS: We have developed a high-throughput qPCR diagnostic assay capable of 
      specific and sensitive quantification of predominant DNA and RNA viruses of 
      cassava in eastern Africa. SIGNIFICANCE AND IMPACT OF THE STUDY: The qPCR methods 
      are a great improvement on the existing methods and can be used for monitoring 
      virus spread as well as for accurate evaluation of the cassava varieties for 
      virus resistance.
CI  - (c) 2016 The Society for Applied Microbiology.
FAU - Otti, G
AU  - Otti G
AD  - Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, UK.
FAU - Bouvaine, S
AU  - Bouvaine S
AD  - Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, UK.
FAU - Kimata, B
AU  - Kimata B
AD  - Naliendele Agricultural Research Institute, Naliendele, Tanzania.
FAU - Mkamillo, G
AU  - Mkamillo G
AD  - Naliendele Agricultural Research Institute, Naliendele, Tanzania.
FAU - Kumar, P L
AU  - Kumar PL
AD  - International Institute of Tropical Agriculture, Ibadan, Nigeria.
FAU - Tomlins, K
AU  - Tomlins K
AD  - Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, UK.
FAU - Maruthi, M N
AU  - Maruthi MN
AD  - Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent, UK.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160311
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
SB  - IM
MH  - Begomovirus/*genetics
MH  - DNA Viruses/genetics
MH  - Manihot/*virology
MH  - Multiplex Polymerase Chain Reaction/methods
MH  - Plant Diseases/*virology
MH  - Potyviridae/*genetics
MH  - RNA Viruses/genetics
MH  - Real-Time Polymerase Chain Reaction/methods
OTO - NOTNLM
OT  - TaqMan assays
OT  - begomovirus
OT  - cassava diseases
OT  - ipomovirus
OT  - real-time PCR
OT  - virus detection and quantification
EDAT- 2016/01/09 06:00
MHDA- 2016/12/27 06:00
CRDT- 2016/01/09 06:00
PHST- 2015/07/28 00:00 [received]
PHST- 2016/01/04 00:00 [revised]
PHST- 2016/01/04 00:00 [accepted]
PHST- 2016/01/09 06:00 [entrez]
PHST- 2016/01/09 06:00 [pubmed]
PHST- 2016/12/27 06:00 [medline]
AID - 10.1111/jam.13043 [doi]
PST - ppublish
SO  - J Appl Microbiol. 2016 May;120(5):1346-56. doi: 10.1111/jam.13043. Epub 2016 Mar 
      11.