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Risk of bias and applicability concerns in the accuracy of Luminex NxTAGG RPP™ for the detection of RSV and influenza viruses in respiratory samples
Source publication
Respiratory syncytial virus (RSV) and influenza viruses are important global causes of morbidity and mortality. We evaluated the diagnostic accuracy of the Luminex NxTAG respiratory pathogen panels (RPPs)™ (index) against other RPPs (comparator) for detection of RSV and influenza viruses. Studies comparing human clinical respiratory samples tested...
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Background
In hospitalized patients, nasopharyngeal (NP) swabs are the most common samples obtained for Respiratory Syncytial Virus (RSV) PCR testing. However, adding sputum is known to increase diagnostic yield, and saliva has been successfully used for viral respiratory infection diagnosis. We sought to compare RSV prevalence detected by PCR test...
Citations
... A key challenge we encountered in our validation efforts was accurate quantitation of the viral cultures. Other groups have reported sensitivity in terms of plaque forming units or 50% tissue culture infectious dose (43)(44)(45)(46). Both of these methods are cumbersome and subjective, and at times yield conflicting results for identical starting cultures in a direct comparison (47,48). ...
... Other methods used for virus quantitation include qPCR and western blots (47). Though faster and easier to use, qPCRs nevertheless pose a similar challenge where two independent assays targeting the same virus can yield differing titers (46,49). We observed this discrepancy firsthand when we used multiple PCR assays in parallel. ...
There is a need in clinical microbiology and public health for assays that enable sensitive and rapid detection of infectious agents. Next Generation Sequencing (NGS) is increasingly used in the fields of oncology and personalized genome medicine but has not gained a wider acceptance for clinical microbiology due to operational and bioinformatics complexity, as well as lower sensitivity compared to agent-specific quantitative polymerase chain reaction (qPCR) assays. VirCapSeq-VERT is a positive selection system for detection, typing, and strain differentiation of both RNA and DNA viruses with sensitivity comparable to qPCR. Here we report the analytical and clinical validation of the VirCapSeq-VERT system for detection of viruses in plasma and nasal secretions that cause systemic and/or respiratory infections.
IMPORTANCE
Broad range assay for accurate and sensitive diagnostics.
... The Luminex® NxTAG RPP is a qualitative molecular test for the identification of nucleic acids from 17 respiratory viruses and 3 bacteria: adenovirus, coronavirus (229E, HKU1, OC43 and NL63), bocavirus, human metapneumovirus, human rhinovirus/enterovirus, influenza virus (A, A/H1-2009, A/H3, B), parainfluenza (1, 2, 3 and 4), RSV, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. The sensitivity and specificity of this test is greater than 95% [14,15]. The Luminex® NxTAG RPP was used according to the instructions in the product leaflet with an approximate duration of 4-5 h. ...
Background and Objective
The clinical relevance of the detection of multiple respiratory viruses in acute bronchiolitis (AB) has not been established. Our goal was to evaluate the effect of viral coinfections on the progression and severity of AB.
Methods
A retrospective observational study was conducted in a tertiary hospital in Spain from September 2012 to March 2020. Infants admitted for AB with at least one respiratory virus identified by molecular diagnostic techniques were included. A comparison was made between single-virus infections and viral coinfections. The evolution and severity of AB were determined based on the days of hospitalization and admission to the pediatric intensive care unit (PICU).
Results
Four hundred forty-five patients were included (58.4% male). The median weight was 5.2 kg (IQR 4.2–6.5), and the median age was 2.5 months (IQR 1.4–4.6). A total of 105 patients (23.6%) were admitted to the PICU. Respiratory syncytial virus (RSV) was the most frequent etiological agent (77.1%). A single virus was detected in 270 patients (60.7%), and viral coinfections were detected in 175 (39.3%), of which 126 (28.3%) had two viruses and 49 (11%) had three or more viruses. Hospital length of stay (LOS) increased in proportion to the number of viruses detected, with a median of 6 days (IQR 4–8) for single infections, 7 days (IQR 4–9) for coinfections with two viruses and 8 days (IQR 5–11) for coinfections with ≥ 3 viruses (p = 0.003). The adjusted Cox regression model showed that the detection of ≥ 3 viruses was an independent risk factor for a longer hospital LOS (HR 0.568, 95% CI 0.410–0.785). No significant association was observed between viral coinfections and the need for PICU admission (OR 1.151; 95% CI 0.737–1.797).
Conclusions
Viral coinfections modified the natural history of AB, prolonging the hospital LOS in proportion to the number of viruses detected without increasing the need for admission to the PICU.
... This model still ensures that sensitivity and specificity are jointly estimated as well as accounting for imperfect reference tests [24] while also allowing for asymmetry in the SROC curve. This model can be seen as a simplification of the random effects model fit in Jullien et al. [25] where the variances of the random effects are zero (i.e.σ θ = 0, and σ α = 0, such thatθ j and α j are equal to � and � , respectively). ...
Background
The incidence of cryptococcosis amongst HIV-negative persons is increasing. Whilst the excellent performance of the CrAg testing in people living with HIV is well described, the diagnostic performance of the CrAg LFA has not been systematically evaluated in HIV-negative cohorts on serum or cerebrospinal fluid.
Methods
We performed a systematic review to characterise the diagnostic performance of IMMY CrAg® LFA in HIV-negative populations on serum and cerebrospinal fluid. A systematic electronic search was performed using Medline, Embase, Global Health, CENTRAL, WoS Science Citation Index, SCOPUS, Africa-Wide Information, LILACS and WHO Global Health Library. Studies were screened and data extracted from eligible studies by two independent reviewers. A fixed effect meta-analysis was used to estimate the diagnostic sensitivity and specificity.
Results
Of 447 records assessed for eligibility, nine studies met our inclusion criteria, including 528 participants overall. Amongst eight studies that evaluated the diagnostic performance of the IMMY CrAg® LFA on serum, the pooled median sensitivity was 96% (95% Credible Interval (CrI) 68–100%) with a pooled specificity estimate of 96% (95%CrI 84–100%). Amongst six studies which evaluated the diagnostic performance of IMMY CrAg® LFA on CSF, the pooled median sensitivity was 99% (95%CrI 95–100%) with a pooled specificity median of 99% (95%CrI 95–100%).
Conclusions
This review demonstrates a high pooled sensitivity and specificity for the IMMY CrAg® LFA in HIV-negative populations, in keeping with findings in HIV-positive individuals. The review was limited by the small number of studies. Further studies using IMMY CrAg® LFA in HIV-negative populations would help to better determine the diagnostic value of this test.
... (1, 2, 3 and 4), RSV, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. Its sensitivity and speci city are greater than 95% [14,15]. The test was performed according to the instructions of the product lea et with an approximate duration of 4 -5 hours. ...
Background and Objective: The clinical relevance of the detection of multiple respiratory viruses in acute bronchiolitis (AB) is not established. Our goal was to evaluate the effect of viral coinfections on the evolution and severity of AB.
Methods: A retrospective observational study was conducted in a tertiary hospital in Spain, from September 2012 to March 2020. Infants admitted for AB, with at least one respiratory virus identified by molecular diagnostic techniques were involved. A comparison was made between single virus infection vs viral coinfections. The evolution and severity of AB were determined based on the days of hospitalization and admission to the Pediatric Intensive Care Unit (PICU).
Results: Four hundred forty-five patients were included (58.4% male). Median weight was 5.2 kg (RIQ 4.2 – 6.5) and age 2.5 months (RIQ 1.4 – 4.6). A total of 105 patients (23.6%) were admitted to PICU. The Respiratory Syncytial virus (RSV) was the most frequent etiological agent (77.1%). A single virus was detected in 270 patients (60.7%) and viral coinfections in 175 (39.3%), of which 126 (28.3%) had two viruses and 49 (11%) had three or more viruses. Hospital length of stay (LOS) increased proportionally to the number of viruses detected, with a median of 6 days (IQR 4 – 8) in single infection, 7 days (IQR 4 – 9) in coinfection by two viruses and 8 days (IQR 5 – 11) in coinfection ≥ 3 viruses, (p=0.003). The adjusted Cox regression model showed that detection of ≥ 3 viruses was an independent risk factor for longer hospital LOS (HR 0.568, 95%CI 0.410 – 0.785). No significant association was observed between viral coinfections and the need for PICU admission (OR 1.151; 95%CI 0.737 – 1.797).
Conclusions: Viral coinfections modify the natural history of AB prolonging the hospital LOS, proportionally to the number of viruses detected, without increasing the need for admission to the PICU.
... [4], [5] Rhinovirus/ Enterovirus (RV/EV) and Respiratory Syncytial Virus (RSV) are the most common causes of respiratory tract infection (RTI) in children. [6] RV/EV can be both asymptomatic and symptomatic with common cold, acute otitis media (AOM), pneumonia, bronchiolitis, recurrent wheezing in children. Young children, the elderly and adults with chronic medical conditions are at the greater risk for severe RSV infections. ...
Background: Acute respiratory infection (ARI) is a major cause of morbidity and mortality worldwide, particularly among children. Viruses are responsible for a large proportion of ARIs, of which Influenza, Parainfluenza, Rhinovirus/Enterovirus (RV/EV) and Respiratory Syncytial Virus (RSV) are the leading cause. Co-infections have a potential role in increase severity of viral illness during COVID-19 pandemic. Human Metapneumovirus, Adenovirus and Bocavirus emerge as single and in co-infection during this period. However, local data on the etiologic diagnosis of ARIs are limited. QIAstat Dx Multiplexed Respiratory panel have recently been added to the diagnostic work-up that can simultaneously detect multiple pathogens of respiratory infections in a single run. Objectives: To determine the prevalence of respiratory viral and atypical bacterial pathogens by rapid multiplex PCR assay, known as the Respiratory Panel, among suspected ARIs patients during COVID-19 pandemic. This study also assesses viral co-infection rates with both COVID-19 and other respiratory viruses. Methods: This retrospective study was conducted at molecular laboratory of SQUARE Hospital Ltd, Dhaka, Bangladesh from March 2020 to October 2022. A total of 1557 respiratory samples were tested by QIAstat Dx Respiratory panel and results were analyzed by software. Results: Of total 1557 tested samples, 458 (29.4%) were positive, 457 respiratory viruses and one Legionella pneumophila. Among total positives, 28.2% were from children and 71.8% from adults with highest numbers found in less than 15 years and more than 60 years of age. Except pandemic virus; SARS-CoV-2 (27.1%), Influenza virus was the most commonly detected pathogen (21.8%) followed by RV/EV (15.9%), RSV(11.6%) and other human Coronaviridae (7.9%). Parainfluenza, Human Metapneumovirus, Adenovirus and Bocavirus were also found, between 2-5%. FLU virus were prevalent among both below15 years and above 60 years but predominance of RV/EV and RSV can be seen in under 15 years. A total of 40 (8.7%) patients were co-infected; 14(35%) with COVID-19 and 26 (65%) between other respiratory viruses, of which the most co-infection was Rhinovirus/Enterovirus plus Bocavirus (15%). Conclusion: Respiratory panel significantly improve etiological diagnosis of multiple respiratory infections because of viruses and atypical bacteria which enhance patient care with more rational antimicrobial use and improving infection control measures during pandemic.
