Diagnostics Rhinovirus Laboratory Detection of Rhinoviruses Jane Kuypers, PhD Senior Research Scientist, Department of Laboratory Medicine, University of Washington


Abstract


Rhinoviruses (RhV) are a frequent cause of the common cold and of more serious illnesses such as otitis media, sinusitis, and asthma exacerbations and more infrequently have been implicated in lower airway infections including pneumonia and exacerbations of chronic obstructive pulmonary disease (COPD) and cystic fibrosis. Laboratory detection of RhV has traditionally been accomplished by culture in tissue cells. Culture methods are laborious, require days to result, and are insensitive. In addition, the recently identified RhV type C strains, which may represent one-third to one-half of RhV genotypes, do not grow in cell culture. RhV detection is currently best achieved using sensitive and specific nucleic acid amplification assays (NAATs). Many NAATs to detect RhV have been developed, including conventional and realtime reverse transcription PCR and nucleic acid sequence-based amplification assays. Most of these assays target the viral 5´ non-coding region, which has more sequence homology among RhV genotypes than other genomic regions. RhV detection is included in several commercially available respiratory virus multiplex PCR panels. Careful design and validation of these NAATs are important to achieve adequate sensitivity and specificity. The recent use of NAAT assays for the detection of RhV in clinical specimens has provided more accurate information about the disease burden and epidemiology of these ubiquitous viruses.


Keywords Rhinoviruses, nucleic acid amplification tests, molecular detection, respiratory virus infections


Disclosure: The author has no conflicts of interest to declare. Received: September 15, 2010 Accepted: November 18, 2010 Citation: US Respiratory Disease, 2010;6:76–80 Correspondence: Jane Kuypers, PhD, 1616 Eastlake Ave E, Suite 320 Seattle, WA 98102. E: kuypers@u.washington.edu


Rhinoviruses (RhV) are important human respiratory pathogens and recently, as a result of testing by more sensitive detection methods, their clinical spectrum of disease has expanded.1–4


binding,22,23 RhV are a common


cause of hospitalization and the most frequently identified respiratory viruses in both upper and lower respiratory tract specimens.1,5 responsible for about two-thirds of common colds2,6 frequent cause of otitis media and sinusitis.4,7,8


of more serious lower airway infections due to RhV have been reported, including bronchiolitis and pneumonia in both immunocompetent and immunocompromised patients5,9–13 asthma and wheezing in children.14,15


RhV-induced wheezing are at high risk for asthma.16,17 They are and are also a In addition, many cases and exacerbations of


Infants who are hospitalized with RhV are also


involved in exacerbations of cystic fibrosis and chronic obstructive pulmonary disease (COPD).18,19


RhV are positive-sense, single-stranded


RhV serotype 87 is now classified as an enterovirus. These original serotypes have also been classified into major and minor receptor groups depending on the human cell surface receptor used for


RNA viruses and members of the picornavirus family, which also includes enteroviruses, parechoviruses, and hepatoviruses. Originally, approximately 100 antigenically distinct serotypes of RhV were identified. Based on RNA sequence data, these serotypes have been classified into two species: RhV A (75 serotypes) and RhV B (25 serotypes).20,21


76


capsid-binding antiviral drugs.24


and into two groups depending on the response to However, the number of RhV strains is


now much larger than the number traditionally identified by serotyping. Recently, new strains in the A species25,26


and multiple novel RhV


genotypes representing a new subtype of RhV, designated RhV C, have been identified using molecular methods,25,27–31 RhV genotypes to over 150.1


bringing the number of RhV C strains have not yet been cultivated


in cell lines, possibly due to a lack of receptors on the cell lines used.1 This article reviews current methods for the laboratory detection of rhinoviruses in clinical specimens.


Traditional Detection Methods


RhV have traditionally been detected in respiratory specimens by their characteristic growth in tissue cell culture and this method is still used by many laboratories. To maximize isolation of RhV serotypes, samples are inoculated onto multiple cell lines, including human diploid fibroblast cell lines (e.g. embryonic lung fibroblast cell lines WI-38 and MRC-5) and the HeLa-1 line overexpressing intercellular adhesion molecule 1 (ICAM-1).8,32


However, detection by culture has several


disadvantages. Cell culture is labor-intensive and requires three to seven days for a result. Specimens must be handled properly and transported to the laboratory quickly so that the viruses remain viable.


© TOUCH BRIEFINGS 2010


Page 1  |  Page 2  |  Page 3  |  Page 4  |  Page 5  |  Page 6  |  Page 7  |  Page 8  |  Page 9  |  Page 10  |  Page 11  |  Page 12  |  Page 13  |  Page 14  |  Page 15  |  Page 16  |  Page 17  |  Page 18  |  Page 19  |  Page 20  |  Page 21  |  Page 22  |  Page 23  |  Page 24  |  Page 25  |  Page 26  |  Page 27  |  Page 28  |  Page 29  |  Page 30  |  Page 31  |  Page 32  |  Page 33  |  Page 34  |  Page 35  |  Page 36  |  Page 37  |  Page 38  |  Page 39  |  Page 40  |  Page 41  |  Page 42  |  Page 43  |  Page 44  |  Page 45  |  Page 46  |  Page 47  |  Page 48  |  Page 49  |  Page 50  |  Page 51  |  Page 52  |  Page 53  |  Page 54  |  Page 55  |  Page 56  |  Page 57  |  Page 58  |  Page 59  |  Page 60  |  Page 61  |  Page 62  |  Page 63  |  Page 64  |  Page 65  |  Page 66  |  Page 67  |  Page 68  |  Page 69  |  Page 70  |  Page 71  |  Page 72  |  Page 73  |  Page 74  |  Page 75  |  Page 76  |  Page 77  |  Page 78  |  Page 79  |  Page 80  |  Page 81  |  Page 82  |  Page 83  |  Page 84