Detection of Apple stem grooving virus from citrus plants by one-step immunocapture reverse transcription polymerase chain reaction (One-Step IC-RT-PCR.

An efficient procedure for the extraction of high-quality RNA from woody plants without the use of phenol, organic solvents, or alcohol precipitation is described. The method employs commercially available spin-column matrices and mitigates the inhibitory effects of plant polysaccharides and polyphenolic compounds commonly observed on subsequent polymerase chain reaction amplification when conventional extraction methods are applied to woody plant species. The method described has been successfully used in the development of highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) techniques for the detection of a number of viruses in their woody hosts. The viruses detected included apple stem grooving capillovirus (ASGV), apple stem pitting virus, Prunus necrotic ringspot ilarvirus (PNRSV), grapevine fanleaf and Arabis mosaic nepoviruses, and grapevine leafroll-associated closterovirus type 3. The method described was equally effective for the extraction of viral RNA from either budwood, leaves, or flower blossoms as determined by the equivalent RT-PCR detection of ASGV and PNRSV from these tissues. Detection of viral RNA in samples of total plant RNA prepared using this method was found to be as sensitive as was previously described for the immunocapture RT-PCR technique.

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Simultaneous detection of six citrus viroids and Apple stem grooving virus from citrus plants by multiplex reverse transcription polymerase chain reaction

Journal of Virological Methods ◽

10.1016/s0166-0934(02)00147-7 ◽

2002 ◽

Vol 106 (2) ◽

pp. 235-239 ◽

Cited By ~ 83

Author(s):

Takao Ito ◽

Hiroyuki Ieki ◽

Katsumi Ozaki

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Simultaneous Detection ◽

Chain Reaction ◽

Apple Stem Grooving Virus ◽

Apple Stem ◽

Citrus Viroids ◽

Polymerase Chain

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A One-Step Reverse Transcription-Polymerase Chain Reaction System for the Simultaneous Detection and Identification of Multiple Tospovirus Infections

Phytopathology ◽

10.1094/phyto-95-0166 ◽

2005 ◽

Vol 95 (2) ◽

pp. 166-171 ◽

Cited By ~ 48

Author(s):

Hiroyuki Uga ◽

Shinya Tsuda

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Simultaneous Detection ◽

Yellow Spot ◽

Rt Pcr ◽

Chain Reaction ◽

Spot Virus ◽

Detection And Identification ◽

One Step ◽

Polymerase Chain

A one-step reverse transcription-polymerase chain reaction (RT-PCR) method has been developed for the simultaneous detection and identification of multiple tospoviruses that infect plants. The RT-PCR system is composed of six primers in a single tube: a universal degenerate primer and five virus species-specific primers. Amplifications resulted in an 848-bp PCR product for Watermelon silver mottle virus, 709-bp for Tomato spotted wilt virus, 589-bp for Impatiens necrotic spot virus, 511-bp for Melon yellow spot virus, and a 459-bp amplicon for Iris yellow spot virus. This system enables the simultaneous detection of at least three types of tospovirus infections, in addition to their species identities, from five possible tospoviruses studied, on the basis of their S RNA combinations. This multiplex RT-PCR system was applied to the detection of tospovirus in ornamental crops cultivated in fields and shows potential for epidemiological studies.

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Pan-filovirus one-step reverse transcription-polymerase chain reaction screening assay

10.1101/579458 ◽

2019 ◽

Cited By ~ 1

Author(s):

Katharina Kopp ◽

Ina Smith ◽

Reuben Klein ◽

Shawn Todd ◽

Glenn A. Marsh ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Disease Ecology ◽

Virus Disease ◽

Virus Species ◽

Rt Pcr ◽

Chain Reaction ◽

Screening Assay ◽

One Step ◽

Polymerase Chain

ABSTRACTFive species within the genera Ebolavirus and Marburgvirus of the family Filoviridae are known to cause severe hemorrhagic fever with high mortality rates in humans and non-human primates. Recent large outbreaks of Ebola virus disease in West Africa (2014 - 2016) and the Democratic Republic of the Congo (2018 - ongoing) have demonstrated the epidemic potential with devastating public health consequences. Several known and novel filovirus species have been found in bats in recent years. However, the role of each virus species in the disease ecology of human disease is still unclear. In particular, the transmission mechanism from potential animal hosts to humans is not known. Therefore, a simple, flexible, cost-effective screening tool for detecting the presence of any (putative) member of the filovirus family in animal samples is needed. In this study, a one-step conventional pan-filovirus RT-PCR assay was developed. The designed universal consensus primers of this screening test target two highly conserved regions of the nucleoprotein (NP) of all currently known filoviruses. The assay was capable of specific amplification of viral RNA of all six primate-pathogenic (human and non-human) filovirus species and resulted in 317 bp long RT-PCR products. This amplicon length renders the assay suitable for flexible application as conventional reverse transcription polymerase chain reaction (RT-PCR) as well as for future use as rapid real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR).

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The occurrence of Apple stem pitting virus and Apple stem grooving virus within field-grown apple cultivars evaluated by RT-PCR

Plant Protection Science ◽

10.17221/3862-pps ◽

2011 ◽

Vol 39 (No. 3) ◽

pp. 88-92 ◽

Cited By ~ 9

Author(s):

J.K. Kundu

Keyword(s):

Polymerase Chain Reaction ◽

Mixed Infection ◽

Rt Pcr ◽

Chain Reaction ◽

Apple Stem Grooving Virus ◽

Apple Stem ◽

Apple Cultivars ◽

Polymerase Chain ◽

Apple Stem Pitting Virus ◽

Stem Pitting

The reverse transcription polymerase chain reaction (RT-PCR) was successfully used to determine the occurrence of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) in field-grown apple cultivars. Both viruses were detected frequently in all 16 tested apple cultivars. As many as 27.86% ASPV-infected and 44% ASGV-infected trees were recorded among a total of 420 tested trees from 15 different orchards. Mixed infection with ASGV and ASPV was recorded in 16.7% of the trees.  

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1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

The Journal of Urology ◽

10.1016/s0022-5347(18)33708-x ◽

2006 ◽

Vol 175 (4S) ◽

pp. 485-486

Author(s):

Sabarinath B. Nair ◽

Christodoulos Pipinikas ◽

Roger Kirby ◽

Nick Carter ◽

Christiane Fenske

Keyword(s):

Prostate Cancer ◽

Polymerase Chain Reaction ◽

Real Time ◽

Molecular Diagnosis ◽

Reverse Transcription ◽

Rt Pcr ◽

Chain Reaction ◽

Polymerase Chain

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Expression of hyaluronan synthase-3 in porcine oviducal epithelium during oestrus

Reproduction Fertility and Development ◽

10.1071/rd02100 ◽

2003 ◽

Vol 15 (2) ◽

pp. 99 ◽

Cited By ~ 22

Author(s):

Paisan Tienthai ◽

Naoko Kimura ◽

Paraskevi Heldin ◽

Eimei Sato ◽

Heriberto Rodriguez-Martinez

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Hyaluronan Synthase ◽

Critical Periods ◽

Rt Pcr ◽

Chain Reaction ◽

Mean Values ◽

Polymerase Chain ◽

Sperm Reservoir ◽

Oviduct Fluid

Hyaluronan (HA) has been related to fertilization and embryo development in the pig. Furthermore, HA is present in pig oviduct fluid and the lining epithelium, particularly of the pre-ovulatory sperm reservoir. Because the mechanisms that regulate HA synthesis have not yet been clarified, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to assess the expression of mRNAs of two HA-synthesizing enzymes (has2 and has3) in the oviduct epithelium (uterotubal junction, isthmus, ampullary–isthmic junction and ampulla segments) of non-inseminated (control) and inseminated (treatment) sows at pre-, peri- and post-ovulatory oestrus. Only has3 mRNA was detected; it was present in all tubal segments of both control and treatment samples. The level of has3 expression did not vary significantly between non-inseminated and inseminated specimens, but there was a tendency (NS) for increased mean values during the peri- and post-ovulatory stages compared with pre-ovulation. It is concluded that has3 is expressed by the porcine endosalpinx epithelium and the levels of expression do not vary during the critical periods of sperm transport and fertilization, despite fluctuating levels of HA in the tubal fluid at corresponding periods.

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