Nested-RT-PCR and multiplex RT-PCR for diagnosis of rhinovirus infection in clinical samples

Xuelian Yu, Reena Ghildyal

Research output: A Conference proceeding or a Chapter in BookChapter

2 Citations (Scopus)

Abstract

Human rhinoviruses (HRVs) are positive-stranded RNA viruses belonging to the Enterovirus genus in the family of Picornaviridae. Identification of the specific strain in HRV disease has been difficult because the traditional serological method is insensitive, labor intensive, and cumbersome. With the fast progress in molecular biological technique, more sensitive and faster molecular methods have been developed, such as polymerase chain reaction (PCR), reverse transcriptase (RT)-PCR, and real-time RT-PCR. To improve the technique for defining the links between illnesses and specific strains of HRV, we developed RT-PCR specific for HRV as routine base. A multiplex RT-PCR that simultaneously identifies 12 respiratory viruses including HRV is also routinely used in our lab. Here we have described the specific steps of methods for identification of HRV from clinical samples, such as sample preparation, isolation of total RNA, nested- RT- PCR for HRV, Seeplex® RV15 ACE Detection method, gel electrophoresis, how to use the QIAxcel® capillary electrophoresis system, and results interpretation.

Original languageEnglish
Title of host publicationRhinoviruses
Subtitle of host publicationMethods and Protocols
EditorsDavid A. Jans, Reena Ghildyal
Place of PublicationNew York, USA
PublisherHumana Press
Chapter2
Pages11-24
Number of pages14
Volume1221
ISBN (Electronic)9781493915712
ISBN (Print)9781493915705
DOIs
Publication statusPublished - 2015

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint Dive into the research topics of 'Nested-RT-PCR and multiplex RT-PCR for diagnosis of rhinovirus infection in clinical samples'. Together they form a unique fingerprint.

  • Cite this

    Yu, X., & Ghildyal, R. (2015). Nested-RT-PCR and multiplex RT-PCR for diagnosis of rhinovirus infection in clinical samples. In D. A. Jans, & R. Ghildyal (Eds.), Rhinoviruses : Methods and Protocols (Vol. 1221, pp. 11-24). (Methods in Molecular Biology). Humana Press. https://doi.org/10.1007/978-1-4939-1571-2_2