Diagnosis Of Respiratory Virus Infections Research Articles

Problems and Methods of Diagnosis of Respiratory Viral Infections of Birds

Problems and Methods of Diagnosis of Respiratory Viral Infections of Birds

Respiratory Viral Infections in Children and Adolescents with Hematological Malignancies.

BackgroundDespite the introduction of a polymerase chain reaction (PCR) test for the diagnosis of respiratory viral infection (RVI), guidance on the application of this test and the management of RVI in immunocompromised children is lacking. This study evaluated the clinical characteristics of RVI and established strategies for the PCR test in children and adolescents with hematological malignancies.MethodsThis study included children and adolescents with underlying hematological malignancies and respiratory symptoms, in whom a multiplex PCR test was performed. Patients in whom RVI was identified and not identified were categorized into Groups I and II, respectively. Group I was sub-divided into patients with upper and lower respiratory infections. The medical records of the enrolled patients were retrospectively reviewed.ResultsA total of 93 respiratory illnesses were included. Group I included 46 (49.5%) cases of RVI, including 31 (67.4%) upper and 15 (32.6%) lower respiratory infections. Rhinovirus (37.0%) was the most common viral pathogen. Significantly more patients in Group I had community-acquired respiratory illnesses (p=0.003) and complained of rhinorrhea (p<0.001) and sputum (p=0.008) than those in Group II. In Group I, significantly more patients with lower respiratory infections had uncontrolled underlying malignancies (p=0.038) and received re-induction or palliative chemotherapy (p=0.006) than those with upper respiratory infections.ConclusionsA multiplex PCR test should be considered for RVI diagnosis in immunocompromised children and adolescents with respiratory symptoms, especially in those with rhinorrhea or sputum prominent over a cough. The early application of the PCR test in patients with uncontrolled underlying malignancies may improve outcomes.

Cellular DNA quantification in respiratory samples for the normalization of viral load: a real need?

BackgroundRespiratory tract infections have an enormous social economic impact, with high incidence of hospitalization and high costs. Adequate specimen collection is the first crucial step for the correct diagnosis of viral respiratory infections. ObjectivesThe present retrospective study aimed: i) to verify the cell yield obtained from sampling the nasal respiratory tract using mid-turbinate flocked swabs; ii) to evaluate the normalization of viral load, based on cell number; and iii) to compare the kinetics of viral infection obtained with normalized vs non-normalized viral load. Study designThe number of cells were quantified by real-time PCR in residual extract of nasal swabs tested for respiratory viruses detection and stored at −80 °C in a universal transport medium (UTM™). ResultsA total of 513 virus-positive and 226 virus-negative samples were analyzed. Overall, a median of 4.42 log10 β2-microgolubin DNA copy number/ml of UTM™ (range 1.17–7.26) was detected. A significantly higher number of cells was observed in virus-positive as compared to virus-negative samples (4.75 vs 3.76; p < 0.001). Viral loads expressed as log10 RNA copies/ml of UTM™ and log10 RNA copies/median number of cells were compared in virus-positive samples and a strict correlation (r = 0.89, p < 0.001) and agreement (R2 = 0.82) were observed. In addition, infection kinetics were compared using the two methods with a follow-up series of eight episodes of viral infection and the mean difference was -0.57 log10 (range −1.99 to 0.40). ConclusionsThe normalization of viral load using cellular load compliments the validation of real-time PCR results in the diagnosis of respiratory viruses but is not strictly needed.

Importance of accuracy of multiplex PCR systems for rapid diagnosis of respiratory virus infection

Importance of accuracy of multiplex PCR systems for rapid diagnosis of respiratory virus infection

Evaluation of a two-stage testing algorithm for the diagnosis of respiratory viral infections

New on-demand multiplex molecular respiratory viral diagnostics offer superior performance although can be expensive and some platforms cannot process multiple specimens simultaneously. We performed a retrospective study reviewing results of patients tested for respiratory viruses following introduction of a two-stage testing algorithm incorporating an initial screen with Sofia® immunoassay then secondary Biofire Filmarray®, and compared to a period when only Filmarray® was used. Of 2976 testing episodes, 1814 underwent initial Sofia® then follow-up FilmArray®. A diagnosis of influenza was made by Sofia® in 282 patients, and by FilmArray® in an additional 163 (median time to result 1.12hours versus 3.46hours, P<0.001). Significantly more patients received their diagnosis within 90minutes in winter despite testing more samples (11.1% versus 3.4%, P<0.001), and approximately $36,000 was saved. An algorithmic approach to respiratory viral diagnosis can combine the advantages of accuracy and speed and be cost saving.

Histopathology of viral infections of the lung.

Lower respiratory viral infections are a significant source of morbidity and mortality worldwide, causing an estimated 3.9 million deaths each year.1 In particular, viruses cause up to 50% of community-acquired pneumonia (CAP) in children and adults, and predispose patients to secondary bacterial pneumonia.1, 2 Those at risk for more severe disease include the very young, very old, immunocompromised, and patients with pre-existing respiratory conditions. Patients with viral pneumonia commonly present with fever, cough, headache, and myalgia. Compared to patients with bacterial or mixed bacterial-viral pneumonia, patients with pure viral pneumonia are less likely to have a productive cough with expectoration.3 Other clinical and radiologic parameters including routine laboratory values and chest radiograph findings are similar between viral and bacterial CAP.3 Overall, the most common causes of viral pneumonia are influenza A virus, influenza B virus, respiratory syncytial virus (RSV), adenovirus and parainfluenza virus.3, 4 With the increased use of nucleic acid amplification testing (NAAT), other viruses including metapneumovirus, rhinovirus and coronaviruses have been identified.5 In the immunocompromised or debilitated host, additional viruses such as cytomegalovirus (CMV), herpes simplex virus (HSV), varicella zoster virus (VZV) are important causes of pneumonia. CMV is the most common cause of viral pneumonia in the immunocompromised host and prophylactic antiviral therapy is commonly administered in patients who are at risk for reactivation.6 RSV is also an important cause of morbidity and mortality in immunosuppressed patients.7 Finally, some vaccine-preventable diseases such as measles pneumonia have increased in prevalence due to low vaccination rates in some populations and increased international travel.8 Laboratory diagnosis of viral pneumonia is usually accomplished through routine testing in the microbiology laboratory using cell culture, antigen detection methods, and increasingly, NAATs such as polymerase chain reaction (PCR).4 Despite the widespread availability of these testing options, viral infections are not always suspected initially and pathologists may therefore play an important role in confirming the presence of infection in histologic and cytologic specimens. In particular, pathologists play an important role in diagnosis of viral respiratory infections at autopsy.7

Rapid Tests for Influenza, Respiratory Syncytial Virus, and Other Respiratory Viruses: A Systematic Review and Meta-analysis.

Rapid diagnosis of respiratory virus infections contributes to patient care. This systematic review evaluates the diagnostic accuracy of rapid tests for the detection of respiratory viruses. We searched Medline and EMBASE for studies evaluating these tests against polymerase chain reaction as the reference standard. Of 179 studies included, 134 evaluated rapid tests for influenza viruses, 32 for respiratory syncytial virus (RSV), and 13 for other respiratory viruses. We used the bivariate random effects model for quantitative meta-analysis of the results. Most tests detected only influenza viruses or RSV. Summary sensitivity and specificity estimates of tests for influenza were 61.1% and 98.9%. For RSV, summary sensitivity was 75.3%, and specificity, 98.7%. We assessed the quality of studies using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) checklist. Because of incomplete reporting, the risk of bias was often unclear. Despite their intended use at the point of care, 26.3% of tests were evaluated in a laboratory setting. Although newly developed tests seem more sensitive, high-quality evaluations of these tests are lacking.

An isothermal, label-free, and rapid one-step RNA amplification/detection assay for diagnosis of respiratory viral infections

Recently, RNA viral infections caused by respiratory viruses, such as influenza, parainfluenza, respiratory syncytial virus, coronavirus, and Middle East respiratory syndrome-coronavirus (MERS-CoV), and Zika virus, are a major public health threats in the world. Although myriads of diagnostic methods based on RNA amplification have been developed in the last decades, they continue to lack speed, sensitivity, and specificity for clinical use. A rapid and accurate diagnostic method is needed for appropriate control, including isolation and treatment of the patients. Here, we report an isothermal, label-free, one-step RNA amplification and detection system, termed as iROAD, for the diagnosis of respiratory diseases. It couples a one-step isothermal RNA amplification method and a bio-optical sensor for simultaneous viral RNA amplification/detection in a label-free and real-time manner. The iROAD assay offers a one-step viral RNA amplification/detection example to rapid analysis (<20min). The detection limit of iROAD assay was found to be 10-times more sensitive than that of real-time reverse transcription-PCR method. We confirmed the clinical utility of the iROAD assay by detecting viral RNAs obtained from 63 human respiratory samples. We envision that the iROAD assay will be useful and potentially adaptable for better diagnosis of emerging infectious diseases including respiratory diseases.

Rapid diagnosis of respiratory viral infections in primary health care

Rapid diagnosis of respiratory viral infections in primary health care

Human metapneumovirus infections in hematopoietic cell transplant recipients and hematologic malignancy patients: A systematic review

Over the past decade, reported incidence of human metapneumovirus (hMPV) has increased owing to the use of molecular assays for diagnosis of respiratory viral infections in cancer patients. The seasonality of these infections, differences in sampling strategies across institutions, and small sample size of published studies make it difficult to appreciate the true incidence and impact of hMPV infections. In this systematic review, we summarized the published data on hMPV infections in hematopoietic cell transplant recipients and patients with hematologic malignancy, focusing on incidence, hMPV-associated lower respiratory tract infection (LRTI), mortality, prevention, and management with ribavirin and/or intravenous immunoglobulins. Although the incidence of hMPV infections and hMPV-associated LRTI in this patient population is similar to respiratory syncytial virus or parainfluenza virus and despite lack of directed antiviral therapy, the mortality rate remains low unless patients develop LRTI. In the absence of vaccine to prevent hMPV, infection control measures are recommended to reduce its burden in cancer patients.

Respiratory RNA Viruses.

Acute upper and lower respiratory infections are a major public health problem and a leading cause of morbidity and mortality worldwide. At greatest risk are young children, the elderly, the chronically ill, and those with suppressed or compromised immune systems. Viruses are the predominant cause of respiratory tract illnesses and include RNA viruses such as respiratory syncytial virus, influenza virus, parainfluenza virus, metapneumovirus, rhinovirus, and coronavirus. Laboratory testing is required for a reliable diagnosis of viral respiratory infections, as a clinical diagnosis can be difficult since signs and symptoms are often overlapping and not specific for any one virus. Recent advances in technology have resulted in the development of newer diagnostic assays that offer great promise for rapid and accurate detection of respiratory viral infections. This chapter emphasizes the fundamental characteristics and clinical importance of the various RNA viruses that cause upper and lower respiratory tract diseases in the immunocompromised host. It highlights the laboratory methods that can be used to make a rapid and definitive diagnosis for the greatest impact on the care and management of ill patients, and the prevention and control of hospital-acquired infections and community outbreaks.

Concordance of nasal swabs and nasopharyngeal swabs in the detection of respiratory viruses by direct immunofluorescence

The most used test for the diagnosis of viral respiratory infection is the detection of viral antigens by direct immunofluorescence (DFA), in samples taken by nasopharyngeal swab (NPS) or aspirate (NPA). It would be desirable to have a less uncomfortable technique to obtain a sample from the patient, but of equal performance. To evaluate the diagnostic agreement between nasal swab (NS) and nasopharyngeal swab (NPS) in the detection of respiratory viruses by DFA and compare the degree of discomfort of both techniques in pediatric patients. Cross-sectional study in children who consulted to a pediatric emergency service with respiratory symptoms. Two samples (NPS and NS) per child were collected. The concordance between the two was determined by Kappa (K) coefficient and the degree of discomfort by a visual pain scale. We obtained 112 samples from 56 children, one by each technique. 82.1% were concordant, K = 0.61 (CI 95%, 0.39-0.83) for the detection of any virus, and K = 0.69 (CI 95%, 0.46-0.92) and K = 0.76 (CI 95%, 0.51-1) for syncytial respiratory virus and influenza A detection, respectively. The degree of discomfort was significantly lower for the NS. There is considerable agreement in the detection of respiratory viruses by DFA between samples obtained by NS and NPS, but not enough to recommend a change in the sampling method in this population.

Respiratory viral infections in infants with clinically suspected pertussis

Objectiveto evaluate the frequency of respiratory viral infections in hospitalized infants with clinical suspicion of pertussis, and to analyze their characteristics at hospital admission and clinical outcomes. Methodsa historical cohort study was performed in a reference service for pertussis, in which the research of respiratory viruses was also a routine for infants hospitalized with respiratory problems. All infants reported as suspected cases of pertussis were included. Tests for Bordetella pertussis (BP) (polymerase chain reaction/culture) and for respiratory viruses (RVs) (immunofluorescence) were performed. Patients who received macrolides before hospitalization were excluded. Clinical data were obtained from medical records. Resultsamong the 67 patients studied, BP tests were positive in 44%, and 26% were positive for RV. There was no etiological identification in 35%, and RV combined with BP was identified in 5%. All patients had similar demographic characteristics. Cough followed by inspiratory stridor or cyanosis was a strong predictor of pertussis, as well as prominent leukocytosis and lymphocytosis. Rhinorrhea and dyspnea were more frequent in viral infections. Macrolides were discontinued in 40% of patients who tested positive for RV and negative for BP. Conclusionthe results suggest that viral infection can be present in hospitalized infants with clinical suspicion of pertussis, and etiological tests may enable a reduction in the use of macrolides in some cases. However, the etiological diagnosis of respiratory virus infection, by itself, does not exclude the possibility of infection with BP.

Longitudinal analysis of leukocyte differentials in peripheral blood of patients with acute respiratory viral infections

BackgroundLeukocyte counts and differentials are commonly acquired in patients with suspected respiratory viral infections and may contribute diagnostic information. However, most published work is limited to a single timepoint at initial presentation to a medical provider, which may correspond to widely varying points in the course of disease. ObjectivesTo examine the temporal development and time-dependent utility of routine leukocyte differentials in the diagnosis of respiratory viral infections. Study designWe analyzed data from recent experimental human challenges with influenza A/H3N2, human rhinovirus (HRV), and respiratory syncytial virus (RSV). Routine clinical lab cell counts and differentials were measured daily from the time period immediately prior to inoculation through the eventual resolution of symptomatic disease. ResultsApproximately 50% of challenged individuals developed symptoms and viral shedding consistent with clinical disease. Subpopulations of WBC showed marked differences between symptomatic and asymptomatic individuals over time, but these changes were much more profound and consistent in influenza infection. Influenza-infected subjects develop both relative lymphopenia and relative monocytosis, both of which closely mirror symptom development in time. A lymphocyte:monocyte ratio of <2 correctly classifies 100% of influenza (but not RSV or HRV) infected subjects at the time of maximal symptoms. ConclusionsLeukocyte differentials may suggest a viral etiology in patients with upper respiratory infection, but are not sufficient to allow differentiation between common viruses. Timing of data acquisition relative to the disease course is a key component in determining the utility of these tests.

A Host-Based RT-PCR Gene Expression Signature to Identify Acute Respiratory Viral Infection

Improved ways to diagnose acute respiratory viral infections could decrease inappropriate antibacterial use and serve as a vital triage mechanism in the event of a potential viral pandemic. Measurement of the host response to infection is an alternative to pathogen-based diagnostic testing and may improve diagnostic accuracy. We have developed a host-based assay with a reverse transcription polymerase chain reaction (RT-PCR) TaqMan low-density array (TLDA) platform for classifying respiratory viral infection. We developed the assay using two cohorts experimentally infected with influenza A H3N2/Wisconsin or influenza A H1N1/Brisbane, and validated the assay in a sample of adults presenting to the emergency department with fever (n = 102) and in healthy volunteers (n = 41). Peripheral blood RNA samples were obtained from individuals who underwent experimental viral challenge or who presented to the emergency department and had microbiologically proven viral respiratory infection or systemic bacterial infection. The selected gene set on the RT-PCR TLDA assay classified participants with experimentally induced influenza H3N2 and H1N1 infection with 100 and 87% accuracy, respectively. We validated this host gene expression signature in a cohort of 102 individuals arriving at the emergency department. The sensitivity of the RT-PCR test was 89% [95% confidence interval (CI), 72 to 98%], and the specificity was 94% (95% CI, 86 to 99%). These results show that RT-PCR-based detection of a host gene expression signature can classify individuals with respiratory viral infection and sets the stage for prospective evaluation of this diagnostic approach in a clinical setting.

Respiratory viral infections in infants with clinically suspected pertussis

Objectiveto evaluate the frequency of respiratory viral infections in hospitalized infants with clinical suspicion of pertussis, and to analyze their characteristics at hospital admission and clinical outcomes. Methodsa historical cohort study was performed in a reference service for pertussis, in which the research of respiratory viruses was also a routine for infants hospitalized with respiratory problems. All infants reported as suspected cases of pertussis were included. Tests for Bordetella pertussis (BP) (polymerase chain reaction/culture) and for respiratory viruses (RVs) (immunofluorescence) were performed. Patients who received macrolides before hospitalization were excluded. Clinical data were obtained from medical records. ResultsAmong the 67 patients studied, BP tests were positive in 44%, and 26% were positive for RV. There was no etiological identification in 35%, and RV combined with BP was identified in 5%. All patients had similar demographic characteristics. Cough followed by inspiratory stridor or cyanosis was a strong predictor of pertussis, as well as prominent leukocytosis and lymphocytosis. Rhinorrhea and dyspnea were more frequent in viral infections. Macrolides were discontinued in 40% of patients who tested positive for RV and negative for BP. Conclusionthe results suggest that viral infection can be present in hospitalized infants with clinical suspicion of pertussis, and etiological tests may enable a reduction in the use of macrolides in some cases. However, the etiological diagnosis of respiratory virus infection, by itself, does not exclude the possibility of infection with BP.

Development of a multiplex one step RT-PCR that detects eighteen respiratory viruses in clinical specimens and comparison with real time RT-PCR

Rapid and accurate diagnosis of viral respiratory infections is crucial for patient management. Multiplex reverse transcriptase polymerase chain reaction (mRT-PCR) is used increasingly to diagnose respiratory infections and has shown to be more sensitive than viral culture and antigen detection. Objective of the present study was to develop a one-step mRT-PCR that could detect 18 respiratory viruses in three sets. The method was compared with real time RT-PCR (rRT-PCR) for its sensitivity and specificity. Clinical specimens from 843 pediatric patients with respiratory symptoms were used in the study. 503 (59.7%) samples were detected positive by mRT-PCR. Of these 462 (54.8%) exhibited presence of a single pathogen and 41 (4.9%) had multiple pathogens. rRT-PCR detected 439 (52.1%) positive samples, where 419 (49.7%) exhibited one virus and 20 (2.4%) showed co-infections. Concordance between mRT-PCR and rRT-PCR was 91.9% and kappa correlation 0.837. Sensitivity and specificity of mRT-PCR were 99.5% and 83.7% while that of rRT-PCR was 86.9% and 99.4% respectively. Rhinovirus (17.2%) was the most frequently detected virus followed by respiratory syncytial virus B (15.4%), H1N1pdm09 (8.54%), parainfluenza virus-3 (5.8%) and metapneumovirus (5.2%). In conclusion, mRT-PCR is a rapid, cost effective, specific and highly sensitive method for detection of respiratory viruses.

Incidence of respiratory viruses in a pediatric population: molecular and epidemiological aspects

Introduction: Respiratory infections are not well defined and the etiology is often unknown. Material and method: four hundred fortynine subjectrs were enrolled in the study; in all patientes there was a suspect of inflammatory diseases of the respiratory tract. At admission, a nasopharyngeal swab was made. A multiplex PCR was performed after extraction and reverse transcription of viral RNA. The amplified fragments were revealed by using an electrophoresis separation. Results: Two hundred and four patients (45.4%) were hospitalized for infection of the upper respiratory tract, 141 (31.4%) for lower respiratory infection and the remaining (23%) for other symptoms. One hundred fiftyseven (35%) patients were positive for human influenza A (H1N1 subtype) and 184 for other respiratory viruses,of which 59 (32%) gave a positive for respiratory syncytial virus, 42 (23%) for rhinovirus, 31 (17%) for parainfluenza virus, 12 (6.5%) for coronavirus, 28 (15%) for adenovirus and 6 (3%) for influenza B (3%) and 6 (3%) for metapneumovirus. The M1 gene sequence of influenza A H1N1 strains from 12 patients had a high identity with that of the reference virus. Conclusion: Furthermore H1N1 and RSV were the main causative agents of acute respiratory infection. A molecular approach provides an accurate and rapid aetiological diagnosis of viral respiratory infections. The molecolar features in the M1 gene suggested that the H1N1 influenza strains circulating in Apulia region had a conserved genetic make up.

Evaluation of serum specific IgM detection in diagnosis of respiratory viral infections in children

The present study was designed to explore the practical application of the rapid etiological diagnosis by detecting specific IgM antibody against common respiratory viruses in children with acute lower respiratory infections (ALRI). Clinical specimens including nasopharyngeal aspirates and serum of acute phase from hospitalized children were collected from 207 infants and children with acute lower respiratory infections from March 2009 to September 2010. Seven common respiratory virus antigens were identified from the collected nasopharyngeal aspirates by direct immunofluorescence assay (DFA). ELISA was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB and PIV, while indirect immunofluorescence assay (IFA) was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB, PIV1, PIV2 and PIV3 in collected acute phase serum. The overall positive rates to detect viral antigen by using DFA, ELISA and IFA was 67.6%, 57.5% and 39.6%, respectively. The consistent rate of ELISA and IFA versus accepted DFA were 21.7% and 31.4%, respectively. The average days from onset of the symptoms to blood sample collection for those with the consistent results by ELISA and DFA were 12.0 d for ADV, 9.6 d for PIV2, 9.5 d for IFV, and 5.3 d for RSV, respectively, and by IFA and DFA were 15.0 d for PIV3, 9.2 d for ADV, and 7.4 d for RSV, respectively. Among all age groups, the consistent rate of serum viral IgM and antigen detections was highest in children younger than 3 years old. Although there were differences between serum IgM antibody and viral antigen detections, specific IgM antibody detection was of value in early and rapid etiological diagnosis of pediatric ALRI, especially for young children. It could provide serologic evidence of respiratory virus infection. The diagnostic rate of pathogen could be improved if it was used in combination with viral antigen diagnostic methods.

134 Validity of a symptom questionnaire for the diagnosis of viral respiratory infection in adults with CF

134 Validity of a symptom questionnaire for the diagnosis of viral respiratory infection in adults with CF