xTAG RVP Research Articles

Cortisol and perceived stress are associated with cytokines levels in patients infected with influenza B virus

Influenza B virus (IBV) causes respiratory infectious disease. Cytokines are important immune mediators during infectious diseases. Cortisol and stress have been related to respiratory infection susceptibility and cytokine regulation. Little is known about systemic cytokines, cortisol, and perceived stress in the early stages of IBV infection. We researched the systemic cytokines and cortisol, as well as the perceived stress and blood cell count in patients infected with IBV. The diagnosis was established using the Luminex xTAG RVP kit and confirmed with qRT-PCR for IBV viral load. The perceived stress was evaluated using the perceived stress scale (PSS-10). Twenty-five plasma cytokines were determined using multiplex immunoassay and cortisol by ELISA. The leukocyte differential count was measured with a standard laboratory protocol. Th1, Th17, and IL-10 cytokines were higher in IBV infected patients (P < 0.05). Leukocytes and neutrophil count negatively correlated with viral load (P < 0.05). Perceived stress had a negative effect on monocyte and systemic cytokines in IBV infected patients (P < 0.05). Cortisol was higher in patients infected with IBV and correlated positively with CCL20 (P < 0.05). Cortisol showed a positive effect on most of the systemic cytokines (P < 0.05). In conclusion, a cytokine pattern was found in IBV infected patients, as well as the possible role of leukocyte counts in the control of IBV. Our results suggest the importance of cortisol and perceived stress on systemic cytokines in patients infected with IBV, but more studies are needed to understand their role in cytokine production in respiratory infectious disease.

Identification of Viruses in Patients With Postviral Olfactory Dysfunction by Multiplex Reverse-Transcription Polymerase Chain Reaction.

Objectives/HypothesisTo investigate causative viruses in patients with postviral olfactory disorders (PVOD).Study DesignCase‐control study.MethodsOne hundred fifty‐one consecutive patients diagnosed with PVOD were enrolled, and samples from 38 patients who visited the doctor within 3 months of symptom onset were collected and analyzed. Thirty‐two individuals who underwent surgery for nasal septal deviation during the same time period were collected as the control group. The Sniffin' Sticks psychophysical olfactory test was used to evaluate olfactory function. Olfactory cleft specimens were collected using nasopharyngeal flocked swabs (COPAN FLOQSwabs). Eighteen viruses were tested for with the Luminex xTAG RVP FAST v2 Assay Kit.ResultsOut of the 38 patients with PVOD, rhinoviruses were detected in 13 patients, and coronavirus OC43 was detected in one patient. The frequency of positive virus detection in the patients with anosmia was higher than in those with hyposmia (58.8% vs. 19.0%, P = 0.018). In control group, rhinovirus was identified in one patient (3.1%). Nasal obstruction was the most common symptom and was experienced by 71.0% of patients.ConclusionsRhinovirus and coronavirus are more commonly identified in PVOD. Our methods represent an approach to screen for viruses that may be involved in PVOD.Level of Evidence4 Laryngoscope, 131:158–164, 2021

Multicenter study of human adenovirus infection in pediatric community-acquired pneumonia in China

Objective: To investigate the predominant genotypes and epidemiological characteristics of human adenovirus (HAdV) in pediatric community-acquired pneumonia (CAP) in China. Methods: This was a repeated cross sectional study. Between November 2014 and November 2016, nasopharyngeal aspirates (NPAs) or throat swabs from each hospitalized pediatric patients diagnosed as CAP in 12 hospitals in Northern and Southern China were collected. Respiratory specimens were screened for 18 respiratory viruses including HAdV by using Luminex xTAG RVP Fast V2 multiplex Assay. Typing of HAdV and analysis for the epidemiological characteristic of HAdV were performed. Results: (1) A total of 2 723 hospitalized pediatric patients with CAP were enrolled in this study and 156 (5.7%, 156/2 723) respiratory specimens were positive for HAdV, and 74 (6.6%, 74/1 128) and 82 (5.1%, 82/1 595) were in Northern and Southern China, respectively. There was no significant difference in the positive detection rate between the Northern and Southern China. (2) In Northern China, the HAdV positive rate of children at the age of <6 months, 6 months-<1 years, 1-<3 years, 3-<5 years and ≥5 years was 5.9%(6/101), 6.7%(7/104), 10.3%(34/331), 4.1%(11/266) and 4.9%(16/326), respectively, and the incidence of HAdV infection peaked in children aged 1-3 years (χ(2)=11.511, P=0.021). While in Southern China the HAdV positive rate of children at the age of <6 months, 6 months-<1 years, 1-<3 years, 3-<5 years and ≥5 years was 2.2% (7/312), 4.6% (12/259), 6.3% (31/494), 7.3% (18/245) and 4.9%(14/285), respectively. There was no significant difference in the positive detection rate among age groups. (3) In 2015, the highest detection rate of HAdV in northern China was 12.5% (25/200) in winter, and in Southern China was 6.7% (35/525) in spring and 5.3% (19/357) in summer. (4) In 108 cases of HAdV positive specimens typing was done and 80 in cases classification was successfully performed.Totally 7 genotypes of HAdV, including HAdV-3 (n=32), HAdV-7 (n=9), HAdV-1 (n=12), HAdV-2 (n=15), HAdV-5 (n=10), HAdV-6 (n=1) and HAdV-4 (n=1), were detected. The predominant HAdV genotypes were HAdV-3 (30.8%, 8/26) and HAdV-7 (26.9%, 7/26) in Northern China, while HAdV-3 (44.4%, 24/54) and HAdV-2 (22.2%, 12/54) were the most prevalent genotypes in Southern China. Conclusions: HAdV is an important viral pathogen in pediatric CAP. The predominant HAdV genotypes and peak seasons of HAdV infections were different between Northern and Southern China. The predominant HAdV genotypes were HAdV-3 and HAdV-7 in Northern China, while HAdV-3 and HAdV-2 in Southern China. The peak season of HAdV infections was winter in Northern China. However, HAdV infections are more common in spring and summer in Southern China.

Cytokines levels in nasal lavage in pediatric patients infected with respiratory viruses

Abstract Respiratory viruses are responsible of acute respiratory infections that affect worldwide population causing morbi-mortality and children younger than 5 years are the most affected population. Cytokines can modulate the immune response but little is known about their levels in nasal mucosa and less if these would correlate with symptoms severity. One hundred twenty-one pediatric patients with clinically diagnosed acute respiratory infections were sequentially enrolled in the Department of Pediatrics of the University Hospital. Throat swabs were performed and nucleic acid was isolated to viral detection by Luminex xTAG RVP kit and qRT-PCR. Nasal lavage were performed in each patients to evaluate cytokines levels by multiplex immunoassay. Disease severity was evaluated according to clinical characteristics on the day of enrollment. The most frequently found viral respiratory infections were hRV, RSV, and hMPV. hMPV load correlated with disease severity (P&amp;lt;0.05). Pro-inflammatory, Th1, Th2 and Th17 cytokines were detected in nasal lavage in those patients, however only pro-inflammatory cytokines were found higher in severe disease. Pro-inflammatory, Th1/Th2/Th17 cytokines correlated with symptoms, mainly with fever (P&amp;lt;0.05). Our findings show that disease severity correlates with viral load and cytokines in nasal lavage may regulate local immune response against respiratory virus.

Epidemiologic and Clinical Characteristics of Coronavirus and Bocavirus Respiratory Infections after Allogeneic Stem Cell Transplantation: A Prospective Single-Center Study

Epidemiologic data about coronaviruses (CoVs) and human bocavirus (HBoV) in the setting of allogeneic hematopoietic stem cell transplantation (allo-HSCT) are scarce. We conducted a prospective longitudinal study on respiratory viral infections (RVIs) in allo-HSCT recipients with respiratory symptoms from December 2013 until June 2016. Respiratory virus in upper and/or lower respiratory tract (URT and LRT) specimens were tested using Luminex xTAG RVP Fast v1 assay. Seventy-nine consecutive allo-HSCT recipients developed a total of 192 virologically documented RVI episodes over 30 months. The median follow-up after RVI was 388 days (range, 5 to 923). CoV or HBoV was detected in 27 of 192 episodes (14%); 18 of 79 recipients (23%) developed a total of 21 CoV RVI episodes, whereas 6 recipients (8%) had 1 HBoV RVI episode each. Fourteen CoV RVI episodes were limited to the URT, whereas 7 affected the LRT. Co-pathogens were detected in 8 (38%) CoV cases. Type OC43 CoV was the dominant type (48%) followed by NL63 (24%), KHU1 (19%), and 229E (9%); the CoV hospitalization rate was 19%, whereas mortality was 5% (1 patient without any other microbiologic documentation). Among the 6 recipients with HBoV (3%), only 1 had LRT involvement and no one died from respiratory failure. In 5 cases (83%) HBoV was detected along with other viral co-pathogens. CoV RVIs are common after allo-HSCT, and in a significant proportion of cases CoV progressed to LRT and showed moderate to severe clinical features. In contrast, HBoV RVIs were rare and mostly presented in the context of co-infections.

Cytokines and miRNAs levels in nasal lavage correlate with symptoms in patients infected with respiratory viruses

Abstract Respiratory viruses are responsible of acute respiratory infections that affect worldwide population causing morbi-mortality. Cytokines and microRNAs can modulate the immune response but little is known in nasal mucosa and less if these would correlate with symptoms severity. We included hundred patients diagnosed with possible viral acute respiratory infections. Patients were sequentially enrolled previous written informed consent was obtained. Virus was detected in pharyngeal exudate by Luminex xTAG RVP kit and qRT-PCR. Nasal lavage were performed in each patients to evaluate microRNAs and cytokines by RT-qPCR, multiplex immunoassay and ELISA, respectively. The symptoms severity was daily monitored throughout seven days by surveys. The respiratory viruses found in these patients were influenza A and B, human rhinovirus, respiratory syncytial virus, human metapneumovirus, parainfluenza, and coronavirus. Viral loads showed no difference between mono-infected patients compared to co-infected; however nasal cytokines and microRNAs profiles differ between these two groups in some cases. Regardless of the viral agent, several nasal microRNAs correlated with symptoms severity. On the other hand, pro-inflammatory and Th2 cytokines correlated positively and negatively with respiratory infection symptoms, respectively. Moreover, correlation between miRNAs and cytokine were found in these patients. Our findings suggest that microRNAs and cytokines in nasal lavage may regulate local immune response against respiratory virus and specific profile may be possible candidates for severity biomarkers.

Evaluation of NxTAG Respiratory Pathogen Panel and Comparison with xTAG Respiratory Viral Panel Fast v2 and Film Array Respiratory Panel for Detecting Respiratory Pathogens in Nasopharyngeal Aspirates and Swine/Avian-Origin Influenza A Subtypes in Culture Isolates.

This study evaluated a new multiplex kit, Luminex NxTAG Respiratory Pathogen Panel, for respiratory pathogens and compared it with xTAG RVP Fast v2 and FilmArray Respiratory Panel using nasopharyngeal aspirate specimens and culture isolates of different swine/avian-origin influenza A subtypes (H2N2, H5N1, H7N9, H5N6, and H9N2). NxTAG RPP gave sensitivity of 95.2%, specificity of 99.6%, PPV of 93.5%, and NPV of 99.7%. NxTAG RPP, xTAG RVP, and FilmArray RP had highly concordant performance among each other for the detection of respiratory pathogens. The mean analytic sensitivity (TCID50/ml) of NxTAG RPP, xTAG RVP, and FilmArray RP for detection of swine/avian-origin influenza A subtype isolates was 0.7, 41.8, and 0.8, respectively. All three multiplex assays correctly typed and genotyped the influenza viruses, except for NxTAG RRP that could not distinguish H3N2 from H3N2v. Further investigation should be performed if H3N2v is suspected to be the cause of disease. Sensitive and specific laboratory diagnosis of all influenza A viruses subtypes is especially essential in certain epidemic regions, such as Southeast Asia. The results of this study should help clinical laboratory professionals to be aware of the different performances of commercially available molecular multiplex RT-PCR assays that are commonly adopted in many clinical diagnostic laboratories.

Detection of Respiratory Viruses from ARTI Patients by xTAG RVP Fast v2 Assay and Conventional Methods

Acute respiratory tract infections (ARTIs) are a major cause of morbidity and mortality in paediatric patients. Therefore, early detection of the viral aetiologies of ARTIs is essential for patient management and infection control. In this study, we evaluated the performance of a new multiplex polymerase chain reaction (PCR) assay (xTAG Respiratory Viral Panel [RVP] Fast v2) in the detection of respiratory viruses by comparing it with that of viral culture and direct immunofluorescence (IF) staining. Nasopharyngeal swab and aspirate samples were collected prospectively from 199 patients who presented with ARTIs at the University Malaya Medical Centre (UMMC) in Kuala Lumpur, Malaysia during a 10-month period. The PCR assay was conducted in parallel with conventional culture and direct IF staining methods. The positive rate of the xTAG RVP Fast v2 assay (78.4%) in detecting respiratory viruses was higher than that of the viral isolation (7.5%) and direct IF (23.1%) methods. Using the xTAG RVP Fast v2 assay, human enterovirus/human rhinovirus (HEV/HRV) was the most frequently detected (46.2%). The xTAG RVP Fast v2 assay revealed mixed infection caused by two or three respiratory viruses in 40 specimens, and these were undetected by the viral isolation and direct IF methods. The xTAG RVP Fast v2 assay was superior to conventional methods in the identification of common respiratory viruses, with higher sensitivity and shorter turnaround times for laboratory results.

Comparing performance of NxTAG RPP, xTAG RVP Fast v2 and FilmArray RP for detecting respiratory pathogens in nasopharyngeal aspirates and swine/avian origin influenza A in culture isolates

Comparing performance of NxTAG RPP, xTAG RVP Fast v2 and FilmArray RP for detecting respiratory pathogens in nasopharyngeal aspirates and swine/avian origin influenza A in culture isolates

Blood microRNAs correlate with plasma cytokines and clinical severity in patients infected with Influenza B and Rhinovirus/Enterovirus.

Abstract Acute respiratory infections (ARI) are the leading causes of infectious disease morbidity and mortality in the world. Two of the most common etiological agents are Influenza B virus (IBV) and Human Rhinovirus (HRV). The IBV is responsible for about 25% of laboratory-documented influenza; meanwhile HRV is responsible for 33% to 50% of common cold cases in adults. Little is known about immunological biomarkers in the early stages of the infection caused by this RNA virus and their possible association with symptoms in those patients. The aim of this study was to identify miRNAs and their correlation with cytokines and clinical severity. Adult patients clinically diagnosed with ARI were sequentially recruited from Medical Service and University Hospital of Universidad Autonoma de Nuevo Leon. The study was conducted with approval of the bioethics committee of the School of Medicine of the Universidad Autonoma de Nuevo Leon. Virus was detected by Luminex xTAG RVP kit, levels of nine miRNAs and twenty six plasma cytokines were analyzed by RT-qPCR and multiplex immunoassay, respectively. The severity of symptoms was daily monitored throughout seven days by surveys. The IBV-positive patients showed lower expression levels of miR-146, miR-16 and increased levels of CCL20 when compared with non-IBV. HRV/Enterovirus(EV)-positive patients showed lower expression levels of miR-132, miR-146 and miR-29. Expression levels of miRNAs and plasma cytokines correlated with symptoms severity in both patient groups. In conclusion, our findings suggest that blood miRNAs and plasma cytokines may regulate systemic immune response against IBV and HRV/EV and many of them may be associated with symptoms severity.

Analysis on xTAG RVP testing results of hospitalized patients with suspected viral pneumonia

To analyze the testing results of hospitalized patients with suspected viral pneumonia using xTAG Respiratory Virus Panel Array (xTAG RVP). 133 hospitalized patients with suspected viral pneumonia from Peking Union Medical College Hospital from January 2013 to December 2014 were divided into two groups, including 42 young and middle-aged patients (17-49 years old) and 91 middle and old-aged patients (50-88 years old). The nucleic acids of sputum or bronchoalveolar lavage fluid (BALF) samples from the two group patients were screened by xTAG RVP and the results were analyzed. Viruses can be detected from 50 out of all 133 samples, and the positive rate was 37.6%. Among the viral positive samples, the proportion of mixed infections was 12.0%. The detection rates of Influenza A virus (FluA) and Rhinovirus (RV) were higher than others (45.0% and 16.7%). The infection rate of young and middle-aged patients was significantly higher than that of the middle and old-aged patients (73.8% vs 31.9%, P<0.001). The viral detection rates of samples from spring, summer, autumn and winter were 25.0%, 6.7%, 20.0% and 38.3%, respectively. The positive rate difference among four seasons was not significant (P=0.257). FluA and RV are dominant respiratory viruses in hospitalized patients with suspected viral pneumonia, and the infection rate of the young and middle-aged patients is higher than the middle and old-aged patients.

Molecular viral epidemiology and clinical characterization of acute febrile respiratory infections in hospitalized children in Taiwan.

Acute respiratory infection (ARI) is a leading cause of morbidity and hospitalization in children. To profile the viruses causing ARI in children admitted to a community‐based hospital in central Taiwan, a cross‐sectional study was conducted on children under 14 years of age that were hospitalized with febrile ARI. Viral etiology was determined using conventional cell culture and a commercial respiratory virus panel fast assay (xTAG RVP), capable of detecting 19 different respiratory viruses and subtype targets. Demographic, clinical, and laboratory data were recorded and analyzed. The RVP fast assay identified at least one respiratory virus in 130 of the 216 specimens examined (60.2%) and rose to 137 (63.4%) by combining the results of cell culture and RVP fast assay. In order of frequency, the etiological agents identified were, rhinovirus/enterovirus (24.6%), respiratory syncytial virus (13.8%), adenovirus (11.5%), parainfluenza virus (9.2%), influenza B (8.4%), influenza A (5.4%), human metapneumovirus (4.6%), human coronavirus (2%), and human bocavirus (2%). Co‐infection did not result in an increase in clinical severity. The RVP assay detected more positive specimens, but failed to detect 6 viruses identified by culture. The viral detection rate for the RVP assay was affected by how many days after admission the samples were taken (P = 0.03). In conclusion, Rhinovirus/enterovirus, respiratory syncytial virus, and adenovirus were prevalent in this study by adopting RVP assay. The viral detection rate is influenced by sampling time, especially if the tests are performed during the first three days of hospitalization. J. Med. Virol. 87:1860–1866, 2015. © 2015 Wiley Periodicals, Inc.

Comparison of the performance of 2 commercial multiplex PCR platforms for detection of respiratory viruses in upper and lower tract respiratory specimens

The performance of the CLART® PneumoVir system with that of the Luminex xTAG RVP Fast v1 assay for detection of most common respiratory viruses in upper and lower tract respiratory specimens (n=183) from unique patients with influenza-like syndrome or lower tract respiratory infection. Nested PCR coupled to automated sequencing was used for resolution of discrepancies. Fully concordant results were obtained for a total of 122 specimens, whereas 56 specimens gave partially (n=21) or fully discordant (n=35) results (Kappa coefficient, 0.62). The overall specificity of the Luminex xTAG RVP Fast v1 assay was slightly higher than that of the CLART® PneumoVir assay for human bocavirus, influenza A virus/H3N2, influenza B virus, human metapneumovirus, and parainfluenza virus, whereas the sensitivity of the latter was higher for most targeted viruses except, notably, for picornaviruses. This was irrespective of either the origin of the respiratory specimen or the age group to which the patients belonged.

Comparison of xTAG respiratory virus panel and Verigene Respiratory Virus Plus for detecting influenza virus and respiratory syncytial virus.

BackgroundNucleic acid amplification tests have allowed simultaneous detection of multiple respiratory viruses.MethodsWe compared the results of a liquid bead array xTAG Respiratory Virus Panel (RVP; (Luminex Corporation, Toronto, Canada) and a solid microarray Verigene Respiratory Virus Plus (RV+; Nanosphere, Northbrook, IL) for the detection of influenza A virus (INF A), influenza B virus (INF B), and respiratory syncytial virus (RSV) in 170 respiratory specimens from hospitalized patients.ResultsOverall, xTAG RVP demonstrated sensitivities and specificities of 97.6 and 100% for INF A, 100 and 99.4% for INF B, and 100 and 100% for RSV, while the Verigene RV+ test sensitivities and specificities were 95.1 and 98.5%, 100.0 and 99.4%, and 97.1 and 100%, respectively. There were no significant differences in the area under the curves between the two assays for each virus (P = 0.364 for INF A, P = 1.000 for INF B, P = 0.317 for RSV). Comparing the results of two assays, discordant results were present mostly due to subtype assignments and identification of coinfections. The detection of viruses was not significantly different (P = 1.000) and the virus/subtype assignment showed good agreement with kappa coefficients of 0.908.ConclusionThe xTAG RVP and Verigene RV+ showed high sensitivities and specificities, and good overall agreement in detection and identification of INF and RSV. These assays can be used in clinical settings for a reliable detection of respiratory viruses found commonly in hospitalized patients.

A case-control study evaluating RT-PCR/ESI-MS technology compared to direct fluorescent antibody and xTAG RVP PCR

Waste nasopharyngeal swabs (N = 244) were evaluated by the reverse-transcriptase polymerase chain reaction/electrospray ionization mass spectrometry PLEX-ID Broad Respiratory Virus Surveillance Kit version 2.5 compared to direct fluorescent antibody and xTAG Respiratory Virus Panel for percent agreement, sensitivity, and specificity. Sensitivity and specificity were 91% (111/122) and 95.1% (116/122), respectively. Sensitivity by virus, except parainfluenza, was 92.9–100%, and specificity was 99–100%.

Detection of respiratory viruses using a multiplex real-time PCR assay in Germany, 2009/10

The aim of this study was to determine the prevalence of respiratory viruses and to prospectively evaluate the performance of the fast-track diagnostics (FTD) respiratory pathogens multiplex PCR assay shortly after the 2009/10 influenza pandemic. Highly sensitive monoplex real-time PCR assays served as references. Discrepant results were further analyzed by the xTAG RVP Fast assay. A total of 369 respiratory samples from children and adults were collected prospectively in Germany from December 2009 until June 2010. The sensitivity and specificity of the FTD assay after resolution of discrepant results was 92.2 % and 99.5 %, respectively. Lowest specificity of the FTD assay was observed for human bocavirus. Multiple detections were recorded in 33/369 (8.9 %) of the samples by monoplex PCR and in 43/369 (11.7 %) using the FTD assay. The most prevalent viruses were respiratory syncytial virus and human metapneumovirus. Only pandemic influenza virus A/H1N1 (2009), and not seasonal influenza virus, was detected. Viruses other than influenza virus accounted for the majority of acute respiratory infections. The FTD assay can be easily implemented in general diagnostic laboratories and facilitate the optimization of patient-management schemes.

A Two-Tube Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Sixteen Human Respiratory Virus Types/Subtypes

There is a need for the development of a rapid and sensitive diagnosis of respiratory viral pathogens. With an intended application in provincial Centers for Diseases Control and Prevention, in this study, we present a two-tube multiplex RT-PCR assay (two-tube assay) using automatic electrophoresis to simultaneously detect sixteen common respiratory viruses. The specificity and the sensitivity of the assay were tested. The assay could detect 20–200 copies per reaction when each viral type was assayed individually, 2000 copies with 9 premixed viral targets in the multiplexed assay in tube 1, and 200 copies with 8 premixed templates in tube 2. A total of 247 specimens were used to evaluate the two-tube assay, and the results were compared with those obtained from the Luminex xTAG RVP Fast assay. The discordant results were confirmed by sequencing or by the Seeplex RV15 ACE detection kit. There were no false positives, but six false negatives occurred with the two-tube assay. In conclusion, the two-tube assay is demonstrated to have great potential for routine surveillance of respiratory virus infection in China.

The development of a GeXP-based multiplex reverse transcription-PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes

BackgroundExisting standard non-molecular diagnostic methods such as viral culture and immunofluorescent (DFA) are time-consuming, labor intensive or limited sensitivity. Several multiplex molecular assays are costly. Therefore, there is a need for the development of a rapid and sensitive diagnosis of respiratory viral pathogens.MethodsA GeXP-based multiplex RT-PCR assay (GeXP assay) was developed to detect simultaneously sixteen different respiratory virus types/subtypes. Seventeen sets of chimeric primers were used to initiate the RT-PCR, and one pair of universal primers was used for the subsequent cycles of the RT-PCR. The specificity of the GeXP assay was examined with positive controls for each virus type/subtype. The sensitivity was evaluated by performing the assay on serial ten-fold dilutions of in vitro-transcribed RNA of all RNA viruses and the plasmids containing the Adv and HBoV target sequence. GeXP assay was further evaluated using 126 clinical specimens and compared with Luminex xTAG RVP Fast assay.ResultsThe GeXP assay achieved a sensitivity of 20–200 copies for a single virus and 1000 copies when all of the 16 pre-mixed viral targets were present. Analyses of 126 clinical specimens using the GeXP assay demonstrated that GeXP assay and the RVP Fast assay were in complete agreement for 109/126 (88.51%) of the specimens. GeXP assay was more sensitive than the RVP Fast assay for the detection of HRV and PIV3, and slightly less sensitive for the detection of HMPV, Adv, RSVB and HBoV. The whole process of the GeXP assay for the detection of 12 samples was completed within 2.5 hours.ConclusionsIn conclusion, the GeXP assay is a rapid, cost-effective, sensitive, specific and high throughput method for the detection of respiratory virus infections.

Comparison of the Luminex xTAG RVP Fast Assay and the Idaho Technology FilmArray RP Assay for Detection of Respiratory Viruses in Pediatric Patients at a Cancer Hospital

Respiratory viruses are increasingly recognized as serious causes of morbidity and mortality in immunocompromised patients. The rapid and sensitive detection of respiratory viruses is essential for the early diagnosis and administration of appropriate antiviral therapy, as well as for the effective implementation of infection control measures. We compared the performance of two commercial assays, xTAG RVP Fast (Luminex Diagnostics, Toronto, Canada) and FilmArray RVP (FA RVP; Idaho Technology, Salt Lake City, UT), in pediatric patients at Memorial Sloan-Kettering Cancer Center. These assays detect the following viruses: respiratory syncytial virus; influenza A and B viruses; parainfluenza viruses 1, 2, 3, and 4; human metapneumovirus; adenovirus; enterovirus-rhinovirus; coronaviruses NL63, HKU1, 229E, and OC43; and bocavirus. We tested a total of 358 respiratory specimens from 173 pediatric patients previously tested by direct fluorescence assay (DFA) and viral culture. The overall detection rate (number of positive specimens/total specimens) for viruses tested by all methods was 24% for DFA/culture, 45% for xTAG RVP Fast, and 51% for FA RVP. The agreement between the two multiplex assays was 84.5%, and the difference in detection rate was statistically significant (P < 0.0001). Overall, the FA RVP assay was more sensitive than the xTAG RVP Fast assay and had a turnaround time of approximately 1 h. The sensitivity, simplicity, and random-access platform make FA RVP an excellent choice for laboratory on-demand service with low to medium volume.

Performance of the Luminex xTAG Respiratory Viral Panel Fast in a clinical laboratory setting

The aim of the study was to develop a real-time RT-PCR for the detection of enteroviruses (EVs) and rhinoviruses (RVs) and to assess the performance of the xTAG RVP Fast assay in comparison to a direct fluorescent assay (DFA), a real-time RT-PCR assay for the detection of respiratory syncytial virus (RSV) and human metapneumovirus (hMPV), and the EV/RV RT-PCR assay developed in this study. The performance of the RVP Fast assay was assessed in the analysis of 373 nasopharyngeal samples. For the viruses of the DFA panel, detection rates of 27.6% and 23.8% were obtained by RVP and DFA, respectively, in analysis of a set of 297 samples collected in 2009-2010. These results show statistically significant superiority of the RVP Fast assay (P=0.049). For RSV, hMPV, EV, and RV, detection rates of 48.0% and 45.2% were achieved by RVP and RT-PCR, respectively. For individual targets, increased detection of EV/RV (P=0.043) and decreased detection of influenza A virus (P=0.004) by RVP in comparison to real-time RT-PCR was observed. The results of the present study imply the need to adjust the InfA component of the RVP Fast assay to also cover the InfA(H1N1) 2009 virus.