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      Non-invasive sample collection for respiratory virus testing by multiplex PCR

      research-article
      a , * , a , b , b , b , c , c , d , e , b , e , f , a
      Journal of Clinical Virology
      Elsevier B.V.
      NPA, nasopharyngeal aspirate, PCR, polymerase chain reaction, RSV, respiratory syncytial virus, PCMC, Primary Children's Medical Center, ED, emergency department, ITI, Idaho Technology, Inc., DFA, direct fluorescent antibody, AV, adenovirus, PIV, parainfluenza virus, hMPV, human metapneumovirus, HRV, human rhinovirus, CV, coronavirus, IV, influenza virus, ILI, influenza-like-illness, Respiratory virus, Viral diagnostics, Molecular diagnostics, Pediatrics, Upper respiratory infection, Polymerase chain reaction

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          Abstract

          Background

          Identifying respiratory pathogens within populations is difficult because invasive sample collection, such as with nasopharyngeal aspirate (NPA), is generally required. PCR technology could allow for non-invasive sampling methods.

          Objective

          Evaluate the utility of non-invasive sample collection using anterior nare swabs and facial tissues for respiratory virus detection by multiplex PCR.

          Study design

          Children aged 1 month–17 years evaluated in a pediatric emergency department for respiratory symptoms had a swab, facial tissue, and NPA sample collected. All samples were tested for respiratory viruses by multiplex PCR. Viral detection rates were calculated for each collection method. Sensitivity and specificity of swabs and facial tissues were calculated using NPA as the gold standard.

          Results

          285 samples from 95 children were evaluated (92 swab-NPA pairs, 91 facial tissue-NPA pairs). 91% of NPA, 82% of swab, and 77% of tissue samples were positive for ≥ 1 virus. Respiratory syncytial virus (RSV) and human rhinovirus (HRV) were most common. Overall, swabs were positive for 74% of virus infections, and facial tissues were positive for 58%. Sensitivity ranged from 17 to 94% for swabs and 33 to 84% for tissues. Sensitivity was highest for RSV (94% swabs and 84% tissues). Specificity was ≥95% for all viruses except HRV for both collection methods.

          Conclusions

          Sensitivity of anterior nare swabs and facial tissues in the detection of respiratory viruses by multiplex PCR varied by virus type. Given its simplicity and specificity, non-invasive sampling for PCR testing may be useful for conducting epidemiologic or surveillance studies in settings where invasive testing is impractical or not feasible.

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          Most cited references34

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          Detection of respiratory viruses by molecular methods.

          Clinical laboratories historically diagnose seven or eight respiratory virus infections using a combination of techniques including enzyme immunoassay, direct fluorescent antibody staining, cell culture, and nucleic acid amplification tests. With the discovery of six new respiratory viruses since 2000, laboratories are faced with the challenge of detecting up to 19 different viruses that cause acute respiratory disease of both the upper and lower respiratory tracts. The application of nucleic acid amplification technology, particularly multiplex PCR coupled with fluidic or fixed microarrays, provides an important new approach for the detection of multiple respiratory viruses in a single test. These multiplex amplification tests provide a sensitive and comprehensive approach for the diagnosis of respiratory tract infections in individual hospitalized patients and the identification of the etiological agent in outbreaks of respiratory tract infection in the community. This review describes the molecular methods used to detect respiratory viruses and discusses the contribution that molecular testing, especially multiplex PCR, has made to our ability to detect respiratory viruses and to increase our understanding of the roles of various viral agents in acute respiratory disease.
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            Rhinovirus transmission within families with children: incidence of symptomatic and asymptomatic infections.

            Rhinoviruses are the most common cause of respiratory tract infections, but the transmission in families has not been studied using sensitive and specific molecular detection methods. Children hospitalized for any infection were screened for rhinoviruses. Eight families with a rhinovirus-positive index child and 16 families with a rhinovirus-negative index child were monitored for 3 weeks for disease symptoms, and the presence and quantity of rhinoviruses in nasal swab samples were determined by quantitative reverse transcription-polymerase chain reaction. Rhinoviruses were further identified by melting temperature and partial sequence analysis. The rates of rhinovirus infection were 1.00 cases per person among the 17 siblings and 0.50 cases per person among the 14 parents of rhinovirus-positive index patients; the rates were 0.54 cases per person among the 24 siblings and 0.23 cases per person among the 30 parents of rhinovirus-negative index patients. Symptomatic infections were associated with an age of <7 years but not with a high copy number of rhinovirus genomes. Virus typing revealed the transmission routes of the viruses and showed that several virus types could circulate in the families simultaneously. Rhinoviruses are frequently transmitted from children to other family members. Most rhinovirus infections in young children are symptomatic, but secondary infections in adults are often asymptomatic. Multiple virus types circulate simultaneously in families.
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              Impact of the rapid diagnosis of influenza on physician decision-making and patient management in the pediatric emergency department: results of a randomized, prospective, controlled trial.

              To determine the impact of the rapid diagnosis of influenza on physician decision-making and patient management, including laboratory tests and radiographs ordered, patient charges associated with these tests, antibiotics/antivirals prescribed, and length of time to patient discharge from the emergency department. Patients aged 2 months to 21 years presenting to an urban children's teaching hospital emergency department were screened for fever and cough, coryza, myalgias, headache, and/or malaise. After obtaining informed consent, patients were randomized to 1 of 2 groups: 1) physician receives (physician aware of) the rapid influenza test result; or 2) physician does not receive (physician unaware of) the result. For patients in the physician aware group, nasopharyngeal swabs were obtained, immediately tested with the FluOIA test for influenza A and B, and the result was placed on the chart before patient evaluation by the attending physician. For the physician unaware group, nasopharyngeal swabs were obtained, stored according to manufacturer's directions, and tested within 24 hours. Results for the physician unaware group were not disclosed to the treating physicians at any time. The 2 resultant influenza-positive groups (aware and unaware) were compared for laboratory and radiograph studies and their associated patient charges, antibiotic/antiviral prescriptions, and length of stay in the emergency department. A total of 418 patients were enrolled, and 391 completed the study. Of these, 202 tested positive for influenza. Comparison of the 96 influenza-positive patients whose physician was aware of the result with the 106 influenza-positive patients whose physician was unaware of the result revealed significant reductions among the former group in: 1) numbers of complete blood counts, blood cultures, urinalyses, urine cultures, and chest radiographs performed; 2) charges associated with these tests; 3) antibiotics prescribed; and 4) length of stay in the emergency department. The number of influenza-positive patients who received prescriptions for antiviral drugs was significantly higher among those whose physician was aware of the result. Physician awareness of a rapid diagnosis of influenza in the pediatric emergency department significantly reduced the number of laboratory tests and radiographs ordered and their associated charges, decreased antibiotic use, increased antiviral use, and decreased length of time to discharge.
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                Author and article information

                Contributors
                Journal
                J Clin Virol
                J. Clin. Virol
                Journal of Clinical Virology
                Elsevier B.V.
                1386-6532
                1873-5967
                19 August 2011
                November 2011
                19 August 2011
                : 52
                : 3
                : 210-214
                Affiliations
                [a ]University of Utah, Department of Pediatrics, Salt Lake City, UT, USA
                [b ]Idaho Technology, Inc., Salt Lake City, UT, USA
                [c ]University of Utah, Department of Internal Medicine, Salt Lake City, UT, USA
                [d ]University of Arizona, College of Public Health, Tucson, AZ, USA
                [e ]Primary Children's Medical Center Microbiology Laboratory, Salt Lake City, UT, USA
                [f ]University of Utah, Department of Pathology, Salt Lake City, UT, USA
                Author notes
                [* ]Corresponding author at: University of Utah Department of Pediatrics, Division of Pediatric Infectious Diseases, 295 Chipeta Way, 2S010, Salt Lake City, UT 84108, USA. Tel.: +1 801 581 6791; fax: +1 801 585 3789. anne.blaschke@ 123456hsc.utah.edu
                Article
                S1386-6532(11)00314-3
                10.1016/j.jcv.2011.07.015
                3196801
                21855405
                a28606fb-379f-4cd2-97c9-24fc1f278761
                Copyright © 2011 Elsevier B.V. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 21 April 2011
                : 13 July 2011
                : 20 July 2011
                Categories
                Article

                Microbiology & Virology
                npa, nasopharyngeal aspirate,pcr, polymerase chain reaction,rsv, respiratory syncytial virus,pcmc, primary children's medical center,ed, emergency department,iti, idaho technology, inc.,dfa, direct fluorescent antibody,av, adenovirus,piv, parainfluenza virus,hmpv, human metapneumovirus,hrv, human rhinovirus,cv, coronavirus,iv, influenza virus,ili, influenza-like-illness,respiratory virus,viral diagnostics,molecular diagnostics,pediatrics,upper respiratory infection,polymerase chain reaction

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