Comparison of four nasal sampling methods for the detection of viral pathogens by RT-PCR—A GA2LEN project

Abstract

The aim of this study was to compare the efficacy and patient discomfort between four techniques for obtaining nasal secretions. Nasal secretions from 58 patients with symptoms of a common cold, from three clinical centers (Amsterdam, Lodz, Oslo), were obtained by four different methods: swab, aspirate, brush, and wash. In each patient all four sampling procedures were performed and patient discomfort was evaluated by a visual discomfort scale (scale 1-5) after each procedure. Single pathogen RT-PCRs for Rhinovirus (RV), Influenza virus and Adenovirus, and multiplex real-time PCR for RV, Enterovirus, Influenza virus, Adenovirus, Respiratory Syncytial Virus (RSV), Parainfluenza virus, Coronavirus, Metapneumovirus, Bocavirus and Parechovirus were performed in all samples. A specific viral cause of respiratory tract infection was determined in 48 patients (83%). In these, the detection rate for any virus was 88% (wash), 79% (aspirate), 77% (swab) and 74% (brush). The degree of discomfort reported was 2.54 for swabs, 2.63 for washes, 2.68 for aspirates and 3.61 for brushings. Nasal washes yielded the highest rate of viral detection without excessive patient discomfort. In contrast, nasal brushes produced the lowest detection rates and demonstrated the highest level of discomfort.

Development and Evaluation of a Flocked Nasal Midturbinate Swab for Self-Collection in Respiratory Virus Infection Diagnostic Testing

“…These findings expand on those of previous studies of nasal sampling in children (1,4,5,8,10) and have implications for the diagnosis and study of respiratory viruses. First, nasal selfsampling was simple, was preferred to staff collection, and may enable testing of more people earlier in the course of their illnesses.…”

Virus shedding and environmental deposition of novel A (H1N1) pandemic influenza virus: interim findings

“…• Nasal swab Although a nasopharyngeal aspirate (NPA) is considered to be the best specimen for detecting influenza A viruses, [53][54][55] this procedure causes more discomfort and is more difficult to perform, particularly in children. Indeed, studies attempting to collect daily NPA samples from subjects have reported problems with subjects' tolerance and compliance with the procedure.…”

Performance of rapid influenza testing in hospitalized children

“…The 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 Reverse transcription of RNA material isolated from nasopharyngeal samples was performed as described previously [12]. RNA was extracted from 50μl of nasal aspirates using Trizol reagent (Invitrogen, CA, USA).…”

“…Four microliters of cDNA were used in the first round PCR reaction mix, while 2μl of the PCR product of the first round were used in the amplification mixture of the second round [14]. Samples were amplified in a PTC-200 DNA Engine thermocycler (MJ Research, MA, USA) with previously described thermocycling conditions [12].…”