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Chemiluminescent detection of potato and pome fruit viroids by digoxigenin-labeled dot blot and tissue blot hybridization.

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A chemiluminescent molecular hybridization protocol was compared to 32P autoradiography for detecting potato spindle tuber viroid (PSTVd) and apple scar skin group viroids (ASSVd). Labeled cRNA probes for PSTVd and ASSVd were synthesized by SP6 RNA polymerase transcription using digoxigenin-11-UTP

Highly sensitive digoxigenin-labelled DNA probe for the detection of potato spindle tuber viroid.

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A molecular probe pSPAv6.2(+), with concatameric insert representing 6.2-times repeated copy of potato spindle tuber viroid (PSTV) RNA, was labelled with digoxigenin and used to detect PSTV by dot-blot hybridization assay. The probe was highly sensitive and specific, detecting as little as 2.5 pg of

Digoxigenin-labelled molecular probe for the simultaneous detection of three potato pathogens: potato spindle tuber viroid (PSTVd), potato virus Y (PVY), and potato leafroll virus (PLRV).

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A molecular probe, p3POT, was constructed of PSTVd, PVY, PLRV cDNA fragments introduced into pUC18 vector. Sequencing of the inserts revealed that cloned fragments covered conservative parts of pathogenic genomes. Dot-blot hybridization of digoxigenin-labelled construct to crude extracts from plants

Digoxigenin-labelled cDNA probes for the detection of potato virus Y in dormant potato tubers.

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An isolate (Y139) of the potato virus Y strain PVYO, common in the potato-growing regions of eastern Canada, was cloned and segments of about 1800 bases from the 3'-end of the PVY-genome were sequenced. These clones were used to prepare molecular detection probes by labelling with digoxigenin. Five

Improved Detection of Potato Leafroll Luteovirus in Leaves and Tubers with a Digoxigenin-Labeled cRNA Probe.

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A digoxigenin-labeled cRNA probe of approximately 2,100 bp was more than 2,000 times more sensitive in detecting potato leafroll virus (PLRV) in leaf extracts of Datura stramonium, Physalis floridana, and potatoes than enzyme-linked immunosorbent assay (ELISA). The limit of detecting PLRV with the

An oligonucleotide array for the identification and differentiation of bacteria pathogenic on potato.

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ABSTRACT Oligonucleotides, 16 to 24 bases long, were selected from the 3' end of the 16S gene and the 16S-23S intergenic spacer regions of bacteria pathogenic on potato, including Clavibacter michiganensis subsp. sepedonicus, Ralstonia solanacearum, and the pectolytic erwinias, including Erwinia

Detection of Potato spindle tuber viroid and Other Related Viroids by a DIG Labelled RNA Probe.

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Viroids can cause diseases of considerable economic importance; in Europe the main concern is with pospiviroids that may affect the tomato and potato industries. Methods for detection are required that are both sensitive and robust. The detection method described here is a probe hybridization method

A mixture of synthetic oligonucleotide probes labeled with biotin for the sensitive detection of potato spindle tuber viroid.

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Five kinds of synthetic oligonucleotide probes labeled with biotin (BIO) were designed for the detection of potato spindle tuber viroid (PSTVd), and their sensitivities were compared with that of a digoxigenin (DIG)- or BIO-labeled cDNA probe. Although each oligonucleotide probe alone was less

Multimeric non-radioactive cRNA probes improve detection of potato spindle tuber viroid (PSTVd).

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Experimental data showed that multimeric, complementary RNA (cRNA) probes, labelled with non-radioactive digoxigenin (DIG), improved sensitivity of detection of the potato spindle tuber viroid (PSTVd) RNA by 2- to 30-fold as compared with corresponding multimeric cDNA probes. The degree of PSTVd

Comparison of nonradioactive cDNA probes for detection of potato spindle tuber viroid by dot-blot hybridization assay.

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Six nonradioactive cDNA probes were compared for their sensitivities for detecting potato spindle tuber viroid (PSTVd) by dot-blot hybridization assay. Three biotinylated PSTVd cDNA probes, labeled by photoactivation with photobiotin, by nick translation or by random priming with biotinylated

First Report of Potato Spindle Tuber Viroid in Costa Rica.

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During 1997 to 1998, symptoms of leaf roll, dwarfism, chlorosis, and occasional leaf necrosis were observed on Solanum tuberosum in several plots in Cartago, the principal potato-production region of Costa Rica. Because of the known association between potato leafroll virus (PLRV) and viroids (1)

Sensitive detection of potato spindle tuber and temperate fruit tree viroids by reverse transcription-polymerase chain reaction-probe capture hybridization.

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A rapid and sensitive assay for the specific detection of plant viroids using reverse transcription-polymerase chain reaction (RT-PCR) -probe capture hybridization (RT-PCR-enzyme-linked immunosorbent assay (ELISA)) was developed. The assay was applied successfully for the detection of potato spindle

Detection of type A, B, E, and F Clostridium botulinum neurotoxins in foods by using an amplified enzyme-linked immunosorbent assay with digoxigenin-labeled antibodies.

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An amplified enzyme-linked immunosorbent assay (ELISA) for the detection of Clostridium botulinum complex neurotoxins was evaluated for its ability to detect these toxins in food. The assay was found to be suitable for detecting type A, B, E, and F botulinum neurotoxins in a variety of food matrices

Comparison of Dig-Labeled PCR, Nested PCR, and ELISA for the Detection of Clavibacter michiganensis subsp. sepedonicus in Field-Grown Potatoes.

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The sensitivity of digoxigenin- (dig-) labeled polymerase chain reaction (PCR) was compared with nested PCR and enzyme-linked immunosorbent assay (ELISA) for the detection of the potato ring rot bacterium, Clavibacter michiganensis subsp. sepedonicus, in seed potatoes and stem tissues. The bacterial

Detection of Phthorimaea operculella granulovirus in vivo and in vitro by a specific DNA probe.

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In analyzing populations of non-infected potato tuber moth (PTM) Phthorimaea operculella, using a total DNA probe from Phthorimaea operculella granulovirus (PhopGV), false positive reactions were obtained indicating homology between cellular and viral DNAs. Using a cloned 2.1 kbp fragment of PhopGV

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