Comparative detection of mild strains of potato spindle tuber viroid from the dormant potato tubers by Return-polyacrylamiee gel electrophoresis and nucleic acid hybridization

1988 ◽  
Vol 31 (1) ◽  
pp. 159-166 ◽  
Author(s):  
R. P. Singh ◽  
A. Boucher
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Potato Spindle Tuber Viroid ◽  
Nucleic Acid Hybridization ◽  
Potato Tubers

A SURVEY OF THE INCIDENCE OF POTATO SPINDLE TUBER VIROID IN PRINCE EDWARD ISLAND USING TWO TESTING METHODS

1988 ◽  
Vol 68 (4) ◽  
pp. 1229-1236 ◽  
Author(s):  
R. P. SINGH ◽  
T-L. DeHAAN ◽  
A. S. JASWAL
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Prince Edward Island ◽  
Potato Crop ◽  
Polyacrylamide Gel Electrophoresis ◽  
Leaf Disc ◽  
Potato Spindle Tuber Viroid ◽  
Polyacrylamide Gel ◽  
Nucleic Acid Hybridization ◽  
Dot Blot

Return-polyacrylamide gel electrophoresis (R-PAGE) and nucleic acid hybridization (dot-blot) were used for the detection of potato spindle tuber viroid (PSTV) from potato leaves. Both methods detected potato plants experimentally infected in the first season or those produced from infected tubers (secondarily infected). PSTV concentration was lower in the first-season infected plants than those in the second. Both methods detected PSTV in a single leaf disc from field-grown infected plants combined with 399 – 499 discs from field-grown healthy plants. The sensitivity of detection by R-PAGE was lower for certain cultivars and increased with the age of plants. About 85 000 leaf samples collected from 123 tablestock fields, 170 seed fields, and 63 cultivars from the Fox Island Elite Seed Farm in Prince Edward Island were found to be free from PSTV infection. Reasons for PSTV absence in the potato crop are discussed.Key words: Diagnosis, dot-blot, return polyacrylamide gel electrophoresis, nucleic acid hybridization, Solanum tuberosum, potato

scholarly journals A Novel Lateral Flow Assay for Rapid and Sensitive Nucleic Acid Detection of Avibacterium paragallinarum

2021 ◽  
Vol 8 ◽  
Author(s):  
Caiyun Huo ◽  
Donghai Li ◽  
Zhenguo Hu ◽  
Guiping Li ◽  
Yanxin Hu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Lateral Flow ◽  
Lateral Flow Assay ◽  
Nucleic Acid Hybridization ◽  
Nucleic Acid Detection ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Infectious Coryza ◽  
Avibacterium Paragallinarum

Avibacterium paragallinarum, the pathogen of infectious coryza, caused a highly contagious respiratory disease that poses a serious threat to chickens. Hence, it is necessary to do diagnostic screening for Av. paragallinarum. Existing technologies have been used for Av. paragallinarum testing, which, however, have some drawbacks such as time consuming and expensive that require well-trained personnel and sophisticated infrastructure, especially when they are limitedly feasible in some places for lack of resources. Nucleic acid hybridization-based lateral flow assay (LFA) is capable of dealing with these drawbacks, which is attributed to the advantages, such low cost, rapid, and simple. However, nucleic acid determination of Av. paragallinarum through LFA method has not been reported so far. In this study, we developed a novel LFA method that employed gold nanoparticle probes to detect amplified Av. paragallinarum dsDNA. Compared with agarose gel electrophoresis, this LFA strip was inexpensive, simple- to- use, and time- saving, which displayed the visual results within 5–8 min. This LFA strip had higher sensitivity that achieved the detection limit of 101 CFU/ml compared with 102 CFU/ml in agarose gel electrophoresis. Besides, great sensitivity was also shown in the LFA strip, and no cross reaction existed for other bacteria. Furthermore, Av. paragallinarum in clinical chickens with infectious coryza were perfectly detected by our established LFA strip. Our study is the first to develop the LFA integrated with amplification and sample preparation techniques for better nucleic acid detection of Av. paragallinarum, which holds great potential for rapid, accurate, and on-site determination methods for early diagnosis of Av. paragallinarum to control further spreading.

Separation of potato spindle tuber viroid ribonucleic acid from Scopolia sinensis into three infectious forms and the purification and oligonucleotide pattern of fraction II RNA. Part IX

1976 ◽  
Vol 54 (7) ◽  
pp. 600-608 ◽  
Author(s):  
R. P. Singh ◽  
J. J. Michniewicz ◽  
S. A. Narang
Keyword(s):  
High Pressure ◽  
Liquid Chromatography ◽  
Nucleic Acid ◽  
Gel Electrophoresis ◽  
Cetyltrimethylammonium Bromide ◽  
Polyacrylamide Gel Electrophoresis ◽  
Potato Spindle Tuber Viroid ◽  
Ribonuclease A ◽  
Reverse Phase ◽  
Polynucleotide Kinase

The existence of three infectious forms of potato spindle tuber viroid (PSTV) RNA from Scopolia sinensis was demonstrated by fractionation with high salt, by reverse phase and high pressure liquid chromatography, and by polyacrylamide gel electrophoresis. Purification of fraction II was achieved by the following steps: extraction of nucleic acid with phenol, precipitation of the RNA with cetyltrimethylammonium bromide, fractionation of the RNA with lithium chloride and isopropanol, and finally gel electrophoresis. A procedure using reverse phase chromatography was developed to obtain 70–90% recovery of RNA from polyacrylamide gels. Purified PSTV fraction II RNA was digested with ribonuclease A and T1 and labelled with [γ-32P]ATP using polynucleotide kinase. The labelled digests were separated by the electrophoresis–homochromatography procedures of Sanger. About 20 and 30 spots were obtained with ribonuclease A and T1, respectively.

Hierarchical Assembly of Viral Nanotemplates with Encoded Microparticles via Nucleic Acid Hybridization

Langmuir ◽  
2008 ◽  
Vol 24 (21) ◽  
pp. 12483-12488 ◽  
Author(s):  
Wui Siew Tan ◽  
Christina L. Lewis ◽  
Nicholas E. Horelik ◽  
Daniel C. Pregibon ◽  
Patrick S. Doyle ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acid Hybridization ◽  
Hierarchical Assembly
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