A multiplex polymerase chain reaction method for reliable, sensitive and simultaneous detection of multiple viruses in citrus trees

Natural Occurrence of mixed infection of Citrus Yellow Mosaic virus (CYMV) and Indian Citrus Ring Spot Virus (ICRSV) and their Detection by Duplex PCR in Sweet Orange

“…A multiplex polymerase chain reaction method was described for reliable sensitive and simultaneous detection of six multiple virus in citrus trees (Roy et al, 2005). Baranwal et al, 2005 have been work out for simultaneous detection of citrus yellow mosaic virus and citrus greening bacterium.…”

PCR bias associated with conserved primer binding sites, used to determine genotype diversity within Citrus tristeza virus populations

“…Information regarding the relative abundance of each genotype within the viral population within a host will yield important information as to genotype selection occurring and competition amongst genotypes. It is therefore important to, not only determine the genotypes present within the CTV population, as achieved by marker analysis (Hilf & Garnsey, 2000;Roy et al, 2005) but also to determine the abundance the known as well as unknown genotypes (Waud et al, 2014). The results of this study confirms that an increased number of nucleotide mismatches between a primer binding site and its conspecific primer leads to decreased ability of primers to amplify the template, especially within a multi-template PCR reaction (Ibarbalz et al, 2014).…”

Recent advances in Citrus psorosis virus

“…Multiplex PCR (mPCR) is a useful technique since different RNA and DNA viruses may infect a single host like citrus. Seven viruses infecting citrus viz., Citrus leaf rugose virus (CLRV), C. psorosis virus (CPsV), C. tatter leaf virus (CTLV), C. tristeza virus (CTV), C. variegation virus (CVV), C. yellow mosaic virus (CYMV), and Indian citrus ringspot virus (ICRSV) belonging to six different virus genera were detected simultaneously by the multiplex PCR (mPCR) assays [57]. Degenerate primers were designed based on the sequences of respective virus isolates.…”

“…The diagnostic technique reduces the risk of contamination, saves time and reduces the cost as compared to other conventional methods for citrus virus detection. Furthermore, the mPCR provides a useful rapid method for detecting multiple virus infections in citrus plants that will aid in the production of virus-free citrus plants for certification programs [57].…”

“…The tools used to detect CPsV in citrus trees are mainly based on using indicator plants and on laboratory tests including enzyme-linked immunosorbent assay (ELISA) [20,33,80]. After CPsV sequences became available, molecular tests based on RT-PCR and dot blot or tissue-print hybridization were also developed for CPsV detection [5,20,26,28,32,55,57,67].…”