Biomaterial Database

One-Step Multiplex Quantitative RT-PCR for the Simultaneous Detection of Viroids and Phytoplasmas. 2019

Ioanna Malandraki, and Christina Varveri, and Nikon Vassilakos

Department of Phytopathology, Laboratory of Virology, Benaki Phytopathological Institute, Athens, Greece.

A one-step multiplex quantitative reverse transcription polymerase chain reaction protocol is described, for the detection in pome trees of Pear blister canker viroid and Apple scar skin viroid, together with universal detection of phytoplasmas. Total nucleic acids extraction is performed according to a modified CTAB protocol and TaqMan MGB probes are used to surpass high genetic variability of viroids. The multiplex real-time assay is at least ten times more sensitive than conventional protocols and its features make it suitable for rapid and massive screening of pome fruit trees phytoplasmas and viroids in certification schemes and surveys.

UI	MeSH Term	Description	Entries
D010935	Plant Diseases	Diseases of plants.	Disease, Plant,Diseases, Plant,Plant Disease
D004269	DNA, Bacterial	Deoxyribonucleic acid that makes up the genetic material of bacteria.	Bacterial DNA
D012367	RNA, Viral	Ribonucleic acid that makes up the genetic material of viruses.	Viral RNA
D012680	Sensitivity and Specificity	Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)	Specificity,Sensitivity,Specificity and Sensitivity
D014772	Viroids	A group of pathogens comprising the smallest known agents of infectious disease. They are unencapsulated and are capable of replicating autonomously in susceptible cells. Positively identified viroids composed of single-stranded RNA have been isolated from higher plants, but the existence of DNA viroids pathogenic to animals is suspected.	Viroid
D045582	Phytoplasma	A genus of minute bacteria in the family ACHOLEPLASMATACEAE that inhabit phloem sieve elements of infected PLANTS and cause symptoms such as yellowing, phyllody, and witches' brooms. Organisms lack a CELL WALL and thus are similar to MYCOPLASMA in animals. They are transmitted by over 100 species of INSECTS especially leafhoppers, planthoppers, and PSYLLIDS.
D060885	Multiplex Polymerase Chain Reaction	Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction.	Multiplex Ligation-Dependent Probe Amplification,Multiplex PCR,Triplex PCR,Triplex Polymerase Chain Reaction,Multiplex Ligation Dependent Probe Amplification,PCR, Multiplex,PCR, Triplex
D020133	Reverse Transcriptase Polymerase Chain Reaction	A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.	Polymerase Chain Reaction, Reverse Transcriptase,Reverse Transcriptase PCR,PCR, Reverse Transcriptase,Transcriptase PCR, Reverse
D027845	Malus	A plant genus in the family ROSACEAE, order Rosales, subclass Rosidae. It is best known as a source of the edible fruit (apple) and is cultivated in temperate climates worldwide.	Apple,Apple Tree,Crab Apple,Malus domestica,Apple Trees,Apple, Crab,Apples,Apples, Crab,Crab Apples,Tree, Apple,Trees, Apple
D031989	Pyrus	A plant genus of the family ROSACEAE known for the edible fruit.	Pears,Pear