scholarly journals Detection of Apple Chlorotic Leaf Spot Virus by Dot-Blot Hybridization.

1991 ◽  
Vol 57 (2) ◽  
pp. 278-282 ◽  
Author(s):  
Yoshiaki KANNO ◽  
Nobuyuki YOSHIKAWA ◽  
Tsuyoshi TAKAHASHI
Keyword(s):  
Leaf Spot ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Spot Virus ◽  
Dot Blot Hybridization ◽  
Chlorotic Leaf

scholarly journals Effect of Thermotherapy on Elimination of Apple Stem Grooving Virus and Apple Chlorotic Leaf Spot Virus for In vitro-cultured Pear Shoot Tips

HortScience ◽  
2006 ◽  
Vol 41 (3) ◽  
pp. 729-732 ◽  
Author(s):  
Liping Wang ◽  
Guoping Wang ◽  
Ni Hong ◽  
Rongrong Tang ◽  
Xiaoyun Deng ◽  
...  
Keyword(s):  
Leaf Spot ◽  
Blot Hybridization ◽  
Shoot Tips ◽  
Dot Blot ◽  
Spot Virus ◽  
Dot Blot Hybridization ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Chlorotic Leaf

Apple stem grooving virus (ASGV) and apple chlorotic leaf spot virus (ACLSV) are two major viruses of pear. In this study, in vitro thermotherapy was carried out at 37°C for 25, 30 and 35 days followed by subculturing of meristem tips of different sizes to eliminate ASGV and ACLSV from pear plants. Virus titers in heat-treated shoot tips were evaluated by ELISA testing of regenerated plants. Results showed that thermotherapy for 35 days significantly decreased the titer of ASGV and ACLSV in cultures regenerated from tips of main and axillary shoots, especially in those from explants 1 mm in length from the tip of meristems. Dot-blot hybridization of biotinylated cDNA probes derived from ACLSV and ASGV was used to detect these viruses in crude tissue extracts of in vitro-grown pear plants. Intense signals were consistently detected in untreated plant samples equivalent to less than 0.5 mg tissue. Comparison of signals from dot-blot hybridization and ELISA absorbance values (A405) confirmed that dot-blot hybridization had a higher sensitivity than PAS-ELISA. Dot-blot hybridization could detect viruses with a titer below the threshold level of ELISA. These results indicate that dot-blot hybridization is a useful tool for large-scale surveys of viruses, which facilitates the production of virus-free propagation materials in certification and sanitation programs. Results of PAS-ELISA and dot-blot hybridization showed that high virus elimination efficiency was achieved by a combination of thermotherapy for 35 days and in vitro culture of 1 mm meristem tips.

scholarly journals RT-PCR and Dot Blot hybridization methods for a universal detection of tospoviruses

2001 ◽  
Vol 26 (2) ◽  
pp. 170-175 ◽  
Author(s):  
MARCELO EIRAS ◽  
RENATO O. RESENDE ◽  
ALEXANDRE A. MISSIAGGIA ◽  
ANTÔNIO C. DE ÁVILA
Keyword(s):  
Blot Hybridization ◽  
Impatiens Necrotic Spot Virus ◽  
Virus Species ◽  
Rt Pcr ◽  
Dot Blot ◽  
Spot Virus ◽  
Dot Blot Hybridization ◽  
Tospovirus Species ◽  
Universal Detection

Transcriptase reverse - polymerase chain reaction (RT-PCR) and dot blot hybridization with digoxigenin-labeled probes were applied for the universal detection of Tospovirus species. The virus species tested were Tomato spotted wilt virus, Tomato chlorotic spot virus, Groundnut ringspot virus, Chrysanthemum stem necrosis virus, Impatiens necrotic spot virus, Zucchini lethal chlorosis virus, Iris yellow spot virus. Primers for PCR amplification were designed to match conserved regions of the tospovirus genome. RT-PCR using distinct primer combinations was unable to simultaneously amplify all tospovirus species and consistently failed to detect ZLCV and IYSV in total RNA extracts. However, all tospovirus species were detected by RT-PCR when viral RNA was used as template. RNA-specific PCR products were used as probes for dot hybridization. This assay with a M probe (directed to the G1/G2 gene) detected at low stringency conditions all Tospovirus species, except IYSV. At low stringency conditions, the L non-radioactive probe detected the seven Tospovirus species in a single assay. This method for broad spectrum detection can be potentially employed in quarantine services for indexing in vitro germplasm.

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

scholarly journals Mapping the RNA-binding domain on the Apple chlorotic leaf spot virus movement protein

2005 ◽  
Vol 86 (1) ◽  
pp. 225-229 ◽  
Author(s):  
Masamichi Isogai ◽  
Nobuyuki Yoshikawa
Keyword(s):  
Leaf Spot ◽  
Rna Binding ◽  
Movement Protein ◽  
Binding Domain ◽  
Sequence Specificity ◽  
Nucleic Acid Binding ◽  
Spot Virus ◽  
Uv Crosslinking ◽  
Chlorotic Leaf ◽  
Rna Binding Domain

The RNA-binding properties of the cell-to-cell movement protein (MP) of Apple chlorotic leaf spot virus were analysed. MP was expressed in Escherichia coli and was used in UV-crosslinking analysis, using a digoxigenin–UTP-labelled RNA probe and gel-retardation analysis. The analyses demonstrated that MP bound cooperatively to single-stranded RNA (ssRNA). When analysed for NaCl dependence of the RNA-binding activity, the majority of the MP could bind ssRNA even in binding buffer with 1 M NaCl. Furthermore, competition binding experiments showed that the MP bound preferentially to ssRNA and single-stranded DNA without sequence specificity. MP deletion mutants were used to identify the RNA-binding domain by UV-crosslinking analysis. Amino acid residues 82–126 and 127–287 potentially contain two independently active, single-stranded nucleic acid-binding domains.

Molecular Characterization and Distribution of Apple Chlorotic Leaf Spot Virus on Apple in China

2013 ◽  
Vol 162 (5) ◽  
pp. 284-290
Author(s):  
Hao Duan ◽  
Zhirui Ji ◽  
Shutong Wang ◽  
Tongle Hu ◽  
Yanan Wang ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Leaf Spot ◽  
Spot Virus ◽  
Chlorotic Leaf

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
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