Establishment and Application of Rapid Diagnosis for Reverse Transcription-Quantitative PCR of Newly Emerging Goose-Origin Nephrotic Astrovirus in China

Abstract

Goose-origin astrovirus (GoAstV), as a newly emerging virus in 2017, is different from previously known astroviruses in the genus Avastrovirus. So far, few studies have focused on the novel virus. Considering the infectious development of astrovirus (AstV), we established a reverse transcription-quantitative PCR (RT-qPCR) assay with a strong specificity to quickly and accurately diagnose GoAstV. Confirmed by clinical application, this method can quickly and accurately detect prevalent GoAstV. The assay is thus convenient for clinical operation and is applicable to the monitoring of GoAstV disease.

Extracellular vesicles from dHL-60 cells as delivery vehicles for diverse therapeutics

“…Although clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) provides robust genome editing, efficient methods for delivering them into target cells are still required. To date, electroporation is the primary choice for delivering CRISPR/Cas9 to target cells 39 , 40 . We examined whether EVs derived from the HL-60 cells could be efficient vehicles for delivering Cas9.…”

Simple and Visible Detection of Novel Astroviruses Causing Fatal Gout in Goslings Using One-Step Reverse Transcription Polymerase Spiral Reaction Method

“…Thus, establishment of a rapid and simple detection method is imperative. Although some PCR-based methods have been successfully used and have proven effective for N-GoAstV detection ( 5 , 7 , 10 , 11 ), the complicated test procedures and expensive equipment required limited their extensive practical use. In the recent years, to circumvent the need for professional laboratory equipment, many detection methods including LAMP and PSR assays were invented for use under isothermal conditions ( 14 – 16 , 19 , 20 ).…”

“…Traditional immunological methods are not ideal for diagnosing novel pathogeny due to the complex preparation required and the need to screen out highly specialized antibodies in the serum. Various molecular-based methods, including reverse transcription-polymerase chain reaction (RT-PCR) ( 5 ) and reverse transcription quantitative PCR (RT-qPCR) ( 10 , 11 ) have been successfully implemented to diagnose N-GoAstV in clinical samples. Despite the fact that these methods offer rapidity, specificity, and sensitivity, they cannot be widely used in the field because of the sophisticated procedures involved and the requirement for expensive instrumentation ( 12 ).…”

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“…Thus, establishment of a rapid and simple detection method is imperative. Although some PCR-based methods have been successfully used and have proven effective for N-GoAstV detection (5,7,10,11), the complicated test procedures and expensive equipment required limited their extensive practical use. In the recent years, to circumvent the need for professional laboratory equipment, many detection methods including LAMP and PSR assays were invented for use under isothermal conditions (14-16, 19, 20).…”

“…Traditional immunological methods are not ideal for diagnosing novel pathogeny due to the complex preparation required and the need to screen out highly specialized antibodies in the serum. Various molecular-based methods, including reverse transcription-polymerase chain reaction (RT-PCR) (5) and reverse transcription quantitative PCR (RT-qPCR) (10,11) have been successfully implemented to diagnose N-GoAstV in clinical samples. Despite the fact that these methods offer rapidity, specificity, and sensitivity, they cannot be widely used in the field because of the sophisticated procedures involved and the requirement for expensive instrumentation (12).…”