Unraveling virus identity by detection of depleted probes with capillary electrophoresis.
With the emergence of new viral infections and pandemics, there is a need to develop faster methods to unravel the virus identities in a large number of clinical samples. This report describes a virus identification method featuring high throughput, high resolution, and high sensitivity detection of viruses. Identification of virus is based on liquid hybridization of different lengths of virus-specific probes to their corresponding viruses. The probes bound to target sequences are removed by a biotin-streptavidin pull-down mechanism and the supernatant is analyzed by capillary electrophoresis. The probes depleted from the sample appear as diminished peaks in the electropherograms and the remaining probes serve as calibrators to align peaks in different capillaries. The virus identities are unraveled by a signal processing and peak detection algorithm developed in-house. Nine viruses were used in the study to demonstrate how the system works to unravel the virus identity in single and double virus infections. With properly designed probes, the system is able to distinguish closely related viruses. The system takes advantage of the high resolution feature of capillary electrophoresis to resolve probes that differ by length. The method may facilitate virus identity screen from more candidate viruses with an automated 4-color DNA sequencer.
The Genomics Research Center, Academia Sinica, Taipei 11529, Taiwan.
Chin-Yu
CY
Wang
Tzu-Hung
TH
Hsiao
Liang-Hui
LH
Chu
Yi-Ling
YL
Lin
Jau-Ling
JL
Huang
Chung-Hsuan
CH
Chen
Konan
K
Peck
D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D019075:Electrophoresis, Capillary; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013997:Time Factors; D014777:Virus Diseases / Q000175:diagnosis / Q000821:virology; D014780:Viruses / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*
D015342:DNA Probes