A Simplified Dot-Blot Hybridization Protocol for Potato spindle tuber viroid Detection in Solanaceae

2021 ◽  
pp. 89-96
Author(s):  
Oxana Kekstidou ◽  
Christina Varveri ◽  
Nikon Vassilakos
Keyword(s):  
Blot Hybridization ◽  
Potato Spindle Tuber Viroid ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Hybridization Protocol

scholarly journals Evidence for Association of a Viroid with Tapping Panel Dryness Syndrome of Rubber (Hevea brasiliensis)

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1155-1155 ◽  
Author(s):  
P. Ramachandran ◽  
S. Mathur ◽  
L. Francis ◽  
A. Varma ◽  
J. Mathew ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Blot Hybridization ◽  
Potato Spindle Tuber Viroid ◽  
Low Molecular Weight ◽  
Total Nucleic Acid ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Low Salt ◽  
Tapping Panel Dryness

Tapping panel dryness (TPD) is one of the most destructive maladies affecting rubber plantations and is becoming a matter of serious concern. Reduced latex yield leading to total drying of the tapping panel is the obvious symptom. The cause of TPD syndrome is unknown but has been mostly attributed to abiotic causes. In India, the high yielding commercial clone RRII 105 is affected by TPD, leading to enormous losses. We have observed that TPD-affected trees show symptoms of bark scaling, cracking, drying, necrotic streaking, and browning of internal bark leading to the decay of internal tissues. Often prominent abnormal bulges on the lower part of tree trunks occur where the first panel begins to dry. Investigations on TPD-affected rubber samples did not reveal the association of fungus, bacterium, virus, or a protozoan. Total nucleic acid extracts purified from leaf and bark tissues of affected samples and analyzed by polyacrylamide gel electrophoresis under denaturing conditions of low salt and high temperature showed the presence of nucleic acids similar in electrophoretic mobility to low molecular weight (LMW) RNA, of ~359 nucleotides such as potato spindle tuber viroid (PSTVd). The LMW nucleic acid detected from TPD-affected samples was found to be RNA based on its sensitivity to RNase and insensitivity to DNase, phenol, and heat treatments. The LMW RNA was purified and cloned in a pUC 19-derived vector by using primers specific to PSTVd (1). The cloned DNA, when random labeled and used as probe reacted specifically to nucleic acid extracts from TPD-affected rubber trees but not from healthy tissue in dot-blot hybridization assays. Based on the above findings, a viroid etiology for TPD syndrome is proposed. Reference: (1) R. A. Owens, A. T. Candresse, and T. O. Diener. Virology 175:238, 1990.

scholarly journals First Report of Potato Spindle Tuber Viroid in Costa Rica

Plant Disease ◽  
1999 ◽  
Vol 83 (11) ◽  
pp. 1072-1072 ◽  
Author(s):  
R. Badilla ◽  
R. Hammond ◽  
C. Rivera
Keyword(s):  
Costa Rica ◽  
Blot Hybridization ◽  
Leaf Roll ◽  
Potato Production ◽  
Potato Spindle Tuber Viroid ◽  
Dot Blot ◽  
First Report ◽  
Production Region ◽  
Dot Blot Hybridization ◽  
Leafroll Virus

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) and previous reports of PLRV in Costa Rica, the presence of potato spindle tuber viroid (PSTVd) was suspected. Leaf samples from 122 symptomatic potato plants, cvs. Atzimba, Floresta, Idiafrit, and Birris, were collected from 10 plots. Total nucleic acids (TNAs) were extracted and purified (2) from collected symptomatic samples and six healthy potato controls. TNAs were spotted on nylon membranes and hybridized to a digoxigenin-labeled DNA probe specific for PSTVd. Of 122 symptomatic plants, 71 were positive for PSTVd based on dot blot hybridization. TNAs from 12 positive potato samples, including at least 1 sample from each cultivar, were used to inoculate Lycopersicum esculentum cv. Super Marmande. Eleven of twelve inoculated tomato plants showed symptoms of dwarfism, leaf deformation, and grayish foliage, often with a dull surface. TNAs were extracted from inoculated tomato and hybridized to the PSTVd probe. All inoculated symptomatic plants were positive for PSTVd based on dot blot hybridization. This is the first report of PSTVd in Costa Rica. References: (1) M. Querci et al. J. Gen. Virol. 78:1207, 1997. (2) W. Villalobos et al. Rev. Biol. Trop. 45:983, 1997.

Feasibility of Qualitative Testing of BCR-ABL and JAK2 V617F in Suspected Myeloproliperative Neoplasm (MPN) Using RT-PCR Reversed Dot Blot Hybridization (RT-PCR RDB)

2019 ◽  
Vol 19 (4) ◽  
pp. 220-227
Author(s):  
Najmiatul Masykura ◽  
Ummu Habibah ◽  
Siti Fatimah Selasih ◽  
Soegiarto Gani ◽  
Cosphiadi Irawan ◽  
...  
Keyword(s):  
Blot Hybridization ◽  
Jak2 V617f ◽  
Rt Pcr ◽  
Dot Blot ◽  
Dot Blot Hybridization

Prevalence of HPV in a Melbourne Female STD Population: Comparison of RNA and DNA Probes in Detecting HPV by Dot Blot Hybridization

1993 ◽  
Vol 4 (3) ◽  
pp. 159-164 ◽  
Author(s):  
A J Borg ◽  
G Medley ◽  
S M Garland
Keyword(s):  
Past History ◽  
Blot Hybridization ◽  
Condylomata Acuminata ◽  
Dot Blot ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Dot Blot Hybridization ◽  
History Of ◽  
Cytological Features ◽  
Dna Types

A total of 377 women, consecutively selected as first attenders to a sexually transmitted diseases clinic in Melbourne, Australia, were examined for overt Condylomata acuminata and were screened for genital HPV DNA types 6, 11, 16, 18, 31, 33 and (35) using 2 dot blot hybridization methods. Overall, there was a 90% positivity correlation between the 2 methods with HPV DNA being detected in 12% of ectocervical samples. Overt warts were found in 15% of the women and HPV DNA was detected at the cervix in 35% with cytology predicting HPV with or without dysplasia in 27%. Thirteen percent had a past history of warts but none on examination and HPV DNA was evident in 16% while 18% had cytological features of HPV. Those with no warts evident and no past history of warts had both HPV DNA and cytological features of HPV in 7%.

Detection of herpes simplex virus and adenovirus DNA by dot blot hybridization using in vitro synthesized RNA transcripts

1986 ◽  
Vol 13 (4) ◽  
pp. 291-299 ◽  
Author(s):  
Volker Schuster ◽  
Bertfried Matz ◽  
Helga Wiegand ◽  
Brigitte Traub ◽  
Dieter Neumann-Haefelin
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Rna Transcripts ◽  
Dot Blot Hybridization ◽  
Simplex Virus

Detection of Cryptosporidium parvum Oocysts on Fresh Vegetables and Herbs Using Antibodies Specific for a Cryptosporidium parvum Viral Antigen

2005 ◽  
Vol 68 (5) ◽  
pp. 1093-1096 ◽  
Author(s):  
K. E. KNIEL ◽  
M. C. JENKINS
Keyword(s):  
Cryptosporidium Parvum ◽  
Viral Antigen ◽  
Blot Hybridization ◽  
Surface Antigens ◽  
Food Samples ◽  
Sensitive Detection ◽  
Dot Blot ◽  
Dot Blot Hybridization ◽  
Fresh Vegetables ◽  
Cryptosporidium Parvum Oocysts

The purpose of this study was to determine if the viral symbiont of Cryptosporidium parvum (CPV) sporozoites could be used as a target for sensitive detection of the parasite in food samples. Polyclonal sera specific to the recombinant viral capsid protein (rCPV40) was used in a dot blot hybridization assay to detect oocysts recovered from green onions and cilantro. Small batches of chopped green onions and cilantro leaves were artificially contaminated with three different concentrations of oocysts: 106, 102, and 101. rCPV40 was superior in detecting oocysts compared with other antibodies directed toward total oocyst protein and oocyst surface antigens. This study provides evidence that CPV is an excellent target for sensitive detection of C. parvum oocysts in foods.

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