Viral Culture Results Research Articles

Accentuating the detection of immunological signatures as a diagnostic tool for monkeypox virus.

Dear Editor, The WHO declared the recent monkeypox surge as a Public Health Emergency of International Concern on 23 July 2022. Rapid measures are being taken in terms of diagnosis, immunization, treatment and management of the disease to prevent another catastrophe. Although monkeypox disease is not proven to be fatal for majority of the people, early diagnosis and appropriate therapeutic regiment has to be followed to curb the transmission of virus. Currently, the predominant diagnostic test performed is the Centers for Disease Control and Prevention (CDC)’s Nonvariola-Orthopox Virus PCR test using samples of skin lesions1. Such invasive methods pose the risk of infection, contamination and reluctant behaviour from patients and healthcare staff while obtaining the sample. This raises the need for a simple, feasible yet accurate mode of detection such as blood or other body fluid investigations. But there is lack of evidence on the use of blood, saliva or other body secretions for detection and confirmation of virus. This article explores the available literature on the detection of immunological signatures in blood as a probable testing method for diagnosing monkeypox virus. The altered kinetics in antiviral antibody response is evident among the confirmed cases of monkeypox disease indicating a significant association between onset of rashes and pattern of rise of immunoglobulins (Igs) in patients’ sera. The viral culture results and PCR finding were found to be were consistent with the seroconversion of Igs. IgM enzyme-linked immunosorbent assay and IgG enzyme-linked immunosorbent assay were thus efficient as diagnostic indicators and phase-of-illness indicators of monkeypox infection2. In addition to this, comparing the serum signatures between monkeypox infected monkey and human cell lines, a characteristic elevation of cluster of differentiation (CD) 40, plasmin and histamine was seen in monkey cell lines, while interferons macrophages and neutrophil-related signalling pathways were predominant in human cell lines3. However, these findings have not been studied in a larger cohort and hence require more intense clinical research to establish these biological constituents as immunological signatures of monkeypox disease. Monkeypox disease has also been strongly associated with early T-cell response and increased inflammatory mediators. An early elevation of CD8+ T cells and reduction in CD4+ T cells is seen in patients with monkeypox virus. This is combined with high expression of CD38 activation marker. This elevated T-cell pattern was found to be normalized within 12–20 days from the onset of symptoms. Inflammatory markers such as interleukins, particularly IL-6, IL-8, IL-16 and tumour necrosis factor are also found to be elevated in these patients. A specific characteristic finding was the detection of Th-1 biased pox-specific T cells in the serum. This antigen specific finding was almost predominant in all of the patients diagnosed with monkeypox virus4,5. Although these studies have provided information about the use of immune markers in detection of monkeypox, the laboratory method of using these immunological signatures for diagnosing monkeypox and disease progression is still at its nascent stage. The detection of immune markers in patient’s serum to identify monkeypox disease will improve the quality and efficacy of testing when compared with conventional skin lesion sampling. Thus, we emphasize on conducting more clinical trials and evidence-based studies to make such immune signatures reliable and accurate for disease detection. Ethical approval Not applicable. Sources of funding None. Authors’ contribution V.P.V.: conceptualization, data curation, writing – original draft preparation. L.P., S.P., J.N.B., U.M.: data curation, writing – original draft preparation. K.M.S.: conceptualization, study design, data curation, writing – original draft preparation, reviewing and editing, visualization and supervision. Conflicts of interest disclosure The authors declare that they have no financial conflict of interest with regard to the content of this report. Research registration unique identifying number (UIN) Not applicable. Guarantor Krishna Mohan Surapaneni.

305. Clinical scoring system to predict viable viral shedding in patients with COVID-19

Abstract Background Centers for Disease Control and Prevention (CDC) recommends 5 to 20 days of isolation for COVID-19 patients depending on symptom duration and severity regardless of genomic PCR results or vaccination history. However, in real clinical practice, more individualized approach is required. We thus developed clinical scoring system to predict viable viral shedding in a given patient by using various factors affecting viable viral shedding. Methods We prospectively enrolled adult patients with SARS-CoV-2 infection admitted to tertiary hospital and day care center between February 2020 and January 2022. The daily dense respiratory sampling (i.e. saliva, sputum, or nasopharyngeal swabs) during the hospital and day care center stay were obtained. Genomic RNA viral load and viral culture were performed for these samples. Clinical predictors of negative viral culture results were identified using survival analysis and multivariable analysis. Results A total of 612 samples from 121 patients of varying degrees of severity were obtained. Of these, 494 (81%) samples were saliva, 63 (10%) were nasopharyngeal swab, and the remaining 55 (9%) were sputum. Of these 612 specimens, 154 (25%) samples revealed positive viral culture results. Univariate and multivariable Cox's time varying proportional hazard model revealed that symptom onset day, viral copy number, disease severity, organ transplant recipient, gender, and vaccination status were independently associated with viral culture results. We thus developed the 5-factor model from -3 to 3 points: viral copy number (-3 to 3 points depending on copy number), disease severity (1 point to moderate to critical diseases), organ transplant recipient (2 points), gender (-1 points to male), and vaccination status (-2 points to fully vaccinated status). The predictive culture-negative rates were calculated through the symptom onset day and the score of the day the sample was collected. Table 1.Prediction scores for negative viral culture resultsTable 2.Predicted Culture-negative Rates Values in the table expressed as percentages. Conclusion Our clinical scoring system can provide objective probability of negative culture results in a given COVID-19 patient with genomic viral load, and appears to be useful to decide de-isolation policy depending on individualized factors associated with viable viral shedding beyond simple symptom-based isolation strategy by CDC. Disclosures All Authors: No reported disclosures.

Correlation of SARS-CoV2 Viral Growth on Cultures, Ct Values, SARS-CoV-2 Variant and Vaccination Status in Asymptomatic, Pre-Symptomatic and Post-Infection Asymptomatic COVID Patients

Abstract Introduction/Objective COVID-19 large scale immunizations have helped control the infectious spread, but breakthrough infections are still observed and some of these patients remain asymptomatic. In this study, we compared the viral growth characteristics with vaccination status in asymptomatic patients. Methods/Case Report We searched our institutional laboratory database for asymptomatic patients infected with delta variant (November-December 2021, n=65) and omicron variant (December 2021, n=68), that were further classified as “truly asymptomatic”, “pre-symptomatic” and “post-infection asymptomatic” based on detailed chart reviews. The patients’ left-over nasopharyngeal swab samples’ cycle threshold values and the recovery of infectious virus in cell culture were compared. Results (if a Case Study enter NA) In the delta cohort, out of 65 asymptomatic patients, 36 were truly asymptomatic, 26 presymptomatic and 3 post infection asymptomatic patients. In the omicron cohort, out of 68 asymptomatic patients, 45 were truly asymptomatic and 23 were presymptomatic patients. The vaccination rate was 59% in truly asymptomatic (delta cohort=56%, omicron cohort=62%), 55% in presymptomatic (delta cohort=50%, omicron cohort=61%) and 67% in post-infection patients (delta cohort only). There was no significant difference in the Ct values of truly asymptomatic (mean Ct=21.08), presymptomatic (mean Ct=20.30) and post-infection asymptomatic patients (mean Ct=20.84) in the combined cohort (p=0.78) and in the delta (p=0.95) or omicron (p=0.45) subcohorts. The Ct values were not correlated with the vaccination status of the patients (vaccinated patients mean Ct=20.56, non-vaccinated patients mean Ct=20.16, p=0.68). Samples with positive viral growth on cultures had significantly lower Ct values (mean Ct= 17.16), compared to samples with negative viral culture results (mean Ct= 24.61) (p<0.0001). The viral growth on culture was not associated with the symptom status, SARS-CoV-2 variant type (delta vs omicron) and vaccination status of the patients in the delta and omicron subcohorts. Conclusion Our findings show that there is no significant correlation between vaccination status and viral loads and culture results in asymptomatic patients.

Clinical scoring system to predict viable viral shedding in patients with COVID-19.

Background: The Centers for Disease Control and Prevention (CDC) recommends 5–10 days of isolation for patients with COVID-19, depending on symptom duration and severity. However, in clinical practice, an individualized approach is required. We thus developed a clinical scoring system to predict viable viral shedding.Methods: We prospectively enrolled adult patients with SARS-CoV-2 infection admitted to a hospital or community isolation facility between February 2020 and January 2022. Daily dense respiratory samples were obtained, and genomic RNA viral load assessment and viral culture were performed. Clinical predictors of negative viral culture results were identified using survival analysis and multivariable analysis.Results: Among 612 samples from 121 patients including 11 immunocompromised patients (5 organ transplant recipients, 5 with hematologic malignancy, and 1 receiving immunosuppressive agents) with varying severity, 154 (25%) revealed positive viral culture results. Multivariable analysis identified symptom onset day, viral copy number, disease severity, organ transplant recipient, and vaccination status as independent predictors of culture-negative rate. We developed a 4-factor predictive model based on viral copy number (-3 to 3 points), disease severity (1 point for moderate to critical disease), organ transplant recipient (2 points), and vaccination status (-2 points for fully vaccinated). Predicted culture-negative rates were calculated through the symptom onset day and the score of the day the sample was collected.Conclusions: Our clinical scoring system can provide the objective probability of a culture-negative state in a patient with COVID-19 and is potentially useful for implementing personalized de-isolation policies beyond the simple symptom-based isolation strategy.

From more testing to smart testing: data-guided SARS-CoV-2 testing choices, the Netherlands, May to September 2020.

BackgroundSARS-CoV-2 RT-PCR assays are more sensitive than rapid antigen detection assays (RDT) and can detect viral RNA even after an individual is no longer infectious. RDT can reduce the time to test and the results might better correlate with infectiousness.AimWe assessed the ability of five RDT to identify infectious COVID-19 cases and systematically recorded the turnaround time of RT-PCR testing.MethodsSensitivity of RDT was determined using a serially diluted SARS-CoV-2 stock with known viral RNA concentration. The probability of detecting infectious virus at a given viral load was calculated using logistic regression of viral RNA concentration and matched culture results of 78 specimens from randomly selected non-hospitalised cases. The probability of each RDT to detect infectious cases was calculated as the sum of the projected probabilities for viral isolation success for every viral RNA load found at the time of diagnosis in 1,739 confirmed non-hospitalised COVID-19 cases.ResultsThe distribution of quantification cycle values and estimated RNA loads for patients reporting to drive-through testing was skewed to high RNA loads. With the most sensitive RDT (Abbott and SD Biosensor), 97.30% (range: 88.65–99.77) of infectious individuals would be detected. This decreased to 92.73% (range: 60.30–99.77) for Coris BioConcept and GenBody, and 75.53% (range: 17.55–99.77) for RapiGEN. Only 32.9% of RT-PCR results were available on the same day as specimen collection.ConclusionThe most sensitive RDT detected infectious COVID-19 cases with high sensitivity and may considerably improve containment through more rapid isolation and contact tracing.

Viral Culture in Hospitalized Congregate Care Patients With Prolonged SARS-CoV-2 Viral RNA Detection

Prolonged detection of SARS-CoV-2 viral RNA has been observed in hospitalized congregate care patients following resolution of clinical symptoms. It is unknown whether patients with persistent PCR positivity pose a risk for COVID-19 transmission. The purpose of this study was to examine the results of serial PCR testing, viral load, and viral culture in patients awaiting discharge prior to a negative PCR test. We sampled 14 patients who were admitted from skilled nursing and/or rehabilitation facilities to a large academic medical center, had clinical signs and symptoms of COVID-19, and had multiple PCR-positive tests separated by at least 14 days. PCR-positive nasopharyngeal swabs were obtained from each patient for viral load quantification and viral culture. The mean age of patients was 72.5 years (55 – 92), with a mean peak SOFA score of 5.6 (1 – 11). Patients were hospitalized for a mean of 37.0 days (25 – 60). RNA was detected by PCR for a mean of 32.9 days (19 – 47). Mean viral load for the first PCR-positive nasopharyngeal swab collected at our hospital was 5.81 genomic copies/mL (2.12 – 9.72). Viral load decreased significantly with days from clinical symptom onset (R = -0.69, 95% CI, -0.80 – -0.55). Four out of 28 samples grew active virus via culture, with no active virus isolates after 2 days of symptom onset. Our viral culture data suggests that persistent PCR positivity may not correlate with infectivity, which has important implications for COVID-19 infection control precautions among older congregate care patients.

Does the SARS-CoV-2 rapid antigen test result correlate with the viral culture result?

Rapid antigen tests (RATs) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have advantages over viral culture in terms of cost and rapidity of testing, but they have low sensitivity. In addition, RATs tend to be negative from approximately 11 days after symptom onset. To determine whether the antigen-negative state indicates a lack of infectiousness, we assessed the association between viral culture and RAT results. Viral culture, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and rapid antigen testing were performed on stored nasopharyngeal samples with threshold cycle values < 30, based on previous RT-qPCR testing. SARS-CoV-2 was isolated by viral culture from nine samples (45%) and one sample (17%) with positive and negative RAT results, respectively. The RAT and viral culture results were both associated with the viral load level and their cutoffs were similar, but the associations were not statistically significant. RAT might be a useful indicator of infectiousness, which can be helpful to control infection. However, further studies with larger sample size are warranted to confirm this observation.

Epidemiologic Characteristics Associated With Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Antigen-Based Test Results, Real-Time Reverse Transcription Polymerase Chain Reaction (rRT-PCR) Cycle Threshold Values, Subgenomic RNA, and Viral Culture Results From University Testing.

BackgroundReal-time reverse transcription polymerase chain reaction (rRT-PCR) and antigen tests are important diagnostics for SARS-CoV-2. Sensitivity of antigen tests has been shown to be lower than that of rRT-PCR; however, data to evaluate epidemiologic characteristics that affect test performance are limited.MethodsPaired mid-turbinate nasal swabs were collected from university students and staff and tested for SARS-CoV-2 using both Quidel Sofia SARS Antigen Fluorescent Immunoassay (FIA) and rRT-PCR assay. Specimens positive by either rRT-PCR or antigen FIA were placed in viral culture and tested for subgenomic RNA (sgRNA). Logistic regression models were used to evaluate characteristics associated with antigen results, rRT-PCR cycle threshold (Ct) values, sgRNA, and viral culture.ResultsAntigen FIA sensitivity was 78.9% and 43.8% among symptomatic and asymptomatic participants respectively. Among rRT-PCR positive participants, negative antigen results were more likely among asymptomatic participants (OR 4.6, CI:1.3-15.4) and less likely among participants reporting nasal congestion (OR 0.1, CI:0.03-0.8). rRT-PCR-positive specimens with higher Ct values (OR 0.5, CI:0.4-0.8) were less likely, and specimens positive for sgRNA (OR 10.2, CI:1.6-65.0) more likely, to yield positive virus isolation. Antigen testing was >90% positive in specimens with Ct values <29. Positive predictive value of antigen test for positive viral culture (57.7%) was similar to that of rRT-PCR (59.3%).ConclusionsSARS-CoV-2 antigen test advantages include low cost, wide availability and rapid turnaround time, making them important screening tests. The performance of antigen tests may vary with patient characteristics, so performance characteristics should be accounted for when designing testing strategies and interpreting results.

Herpes Simplex Infection During Pregnancy, Results of a Tertiary Referral Center in Turkey.

The present study aimed to determine herpes simplex virus (HSV) infection during pregnancy, serologic status of symptomatic patients, and the frequency of PCR and culture positivity and neonatal disease in an obstetric population. Sera of 1,690 women with genital lesions were tested for HSV IgM and/or HSV IgG between 2008 and 2017. All the patients were evaluated by PCR and viral culture to demonstrate genital herpes infection. The frequency of HSV IgG and IgM seropositivity was 73.8% (1245/1685) and 28.6% (484/1690). The number with a positive viral culture was 288, whereas positive PCR was 305. Five infants had localized skin lesions of HSV infection at the time of birth. This study shows the serologic status, viral culture and PCR testing results of pregnant women who had a symptomatic genital HSV infection and efficacy of management strategies to prevent neonatal herpes infection.

Use of ATP Readings to Predict a Successful Hygiene Intervention in the Workplace to Reduce the Spread of Viruses on Fomites.

The purpose of this study was to validate the use of adenosine triphosphate (ATP) for evaluating hygiene intervention effectiveness in reducing viral dissemination in an office environment. The bacterial virus MS-2 was used to evaluate two scenarios, one where the hand of an individual was contaminated and another where a fomite was contaminated. MS-2 was selected as a model because its shape and size are similar to many human pathogenic viruses. Two separate experiments were conducted, one in which the entrance door push plate was inoculated and the other in which the hand of one selected employee was inoculated. In both scenarios, 54 selected surfaces in the office were tested to assess the dissemination of the virus within the office. Associated surface contamination was also measured employing an ATP meter. More than half of the tested hands and surfaces in the office were contaminated with MS-2 within 4h. Next, an intervention was conducted, and each scenario was repeated. Half of the participating employees were provided hand sanitizer, facial tissues, and disinfecting wipes, and were instructed in their use. A significant (p<0.05) reduction was observed in the number of surfaces contaminated with virus. This reduction in viral spread was evident from the results of both viral culture and the surface ATP measurements, although there was no direct correlation between ATP measurements with respect to viral concentration. Although ATP does not measure viruses, these results demonstrate that ATP measurements could be useful for evaluating the effectiveness of hygiene interventions aimed at preventing viral spread in the workplace.

Analysis of clinical information and reverse transcriptase-polymerase chain reaction for early diagnosis of enteroviral meningitis

PurposeMeningitis is among the most common infections affecting the central nervous system. It can be difficult to determine the exact pathogen responsible for the infection and patients are often treated with empiric antibiotics. This study was conducted to identify the most common clinical characteristics of enteroviral meningitis in children and evaluate the diagnostic efficacy of reverse transcriptase-polymerase chain reaction (RT-PCR) for early detection of an enterovirus.MethodsWe analyzed the medical records of children admitted to Korea University Medical Center and diagnosed with meningitis on the basis of cerebrospinal fluid (CSF) analysis and RT-PCR from CSF and other samples from January 2010 to August 2013.ResultsA total of 333 patients were enrolled and classified into four groups based on diagnosis: enteroviral meningitis (n=110), bacterial meningitis (n=23), other viral meningitis (n=36), and unknown etiology (n=164). Patients with bacterial meningitis were younger than those in the other groups (P<0.001). Pleocytosis in CSF was similar across all groups. Of patients in the enteroviral meningitis group, 92.7% were diagnosed based on RT-PCR findings. Mean length of hospital stay for patients with enteroviral meningitis was 6.08 days, which was significantly shorter than that for patients with meningitis of bacterial etiology (19.73 days, P<0.001).ConclusionDiagnosis of enteroviral meningitis before viral culture results are available is possible using RT-PCR. Accurate diagnosis reduces the length of hospital stay and helps to avoid unnecessary empiric antibiotic treatment.

Successful treatment of eczema coxsackium with wet wrap therapy and low-dose topical corticosteroid

Successful treatment of eczema coxsackium with wet wrap therapy and low-dose topical corticosteroid

Clinical and Laboratory Characteristics of Disseminated Herpes Simplex Virus Infection in Neonates

Disseminated herpes simplex virus (HSV) infection is the most fulminant type of neonatal HSV infection and has the highest mortality. Early diagnosis and treatment are essential for patient survival. We describe the clinical presentation, laboratory characteristics, and outcomes of neonates with disseminated HSV infection at our institution. A retrospective review of electronic medical records from 2006 to 2013 was performed. Only neonates with disseminated HSV infection, confirmed by using polymerase chain reaction or viral culture results, were included. Twenty-two cases were identified; the age range was 1 to 14 days. The majority of patients did not have a maternal history of HSV or a history of maternal fever at delivery. Eleven of the patients were delivered by cesarean delivery, and 3 of these patients did not have prolonged rupture of membranes. Neonatal fever, the most common historical characteristic, was present in only one-half of the patients. Pneumonia and respiratory distress were present in one-half of the patients. Serum aspartate aminotransferase and alanine aminotransferase levels were elevated in most, but not all, patients. The blood HSV polymerase chain reaction was positive in all patients tested. Of the 22 study patients, 16 survived and 6 died. The majority of the patients who died had respiratory disease and a delay in the initiation of acyclovir therapy. Disseminated HSV infection in neonates can be challenging to diagnose and is associated with high mortality. Clinicians must strongly consider this diagnosis, test the blood for HSV polymerase chain reaction, and initiate early treatment in the appropriate clinical scenarios.

The accuracy of Influenza A (H1N1) “swine flu” laboratory testing: A systematic review of diagnostic test accuracy

Background Influenza A (H1N1) recently became pandemic, highlighting the need for a cheap and accurate diagnostic test to diagnose this virus in a clinically relevant timeframe. The current reference standard (viral culture) requires a significant degree of technical expertise, laboratory time, resources and can take up to ten days to obtain a result, during which time there could be a significant spread of infection. Objectives To obtain summary estimates of the diagnostic accuracy of currently available laboratory tests compared to viral culture for the diagnosis of Influenza A (H1N1) from respiratory samples. Inclusion criteria Types of participants Patients presenting with influenza-like illnesses and who underwent both an index and reference test specific for influenza A (H1N1). Focus of the review The diagnostic test accuracy of tests for laboratory tests for Influenza A (H1N1) Types of studies Diagnostic test accuracy studies that compared the diagnostic accuracy of any laboratory test (index test) compared to viral culture as the reference test were considered for inclusion. Search strategy Diagnostic tests are still poorly indexed by major databases; therefore the search strategy was deliberately very broad and was conducted across 14 major databases during May 2010. Methodological quality Methodological quality was determined using the QUADAS checklist. Overall the methodological quality of the studies was moderate, however all suffered from incomplete reporting. Data collection Data was collected to determine the accuracy of the index tests and completeness of the conduct and reporting. Patients were identified as being either H1N1 positive or negative on the basis of the viral culture results. The index test results were then compared for sensitivity and specificity. Data synthesis Meta analysis was not appropriate as the included studies were heterogeneous, therefore data was synthesized in narrative summary. Results The search identified 3843 studies. Of these 56 full text articles were retrieved for further analysis. Twenty nine articles were critically appraised, with 24 being excluded on the basis of incongruence with the review objective or for containing insufficient detail. The remaining five cross sectional studies examined the diagnostic accuracy of polymerase chain reaction (PCR), and were included. Two studies reported accuracy measures and when reported, the sensitivity and specificity of PCR was high. Conclusions Although unable to generate a summary estimate of the overall accuracy of PCR, the available evidence suggests that PCR using respiratory specimens appears to be highly sensitive and specific at identifying patients with H1N1. Implications for practice When reported, PCR had high test sensitivity and specificity compared with viral culture, suggesting that patients were accurately being diagnosed by this method and in a much shorter timeframe. Caution should be taken interpreting these results, as this is based on the findings from two included studies Implications for research This systematic review highlights the need for better reporting and indexing of studies that examine the diagnostic accuracy of laboratory tests in general, and H1N1 specifically. This rapidly expanding area of research needs structure and consistency in order for its findings to be useful to health care clinicians and decision makers.

Azathioprine-induced fever in autoimmune hepatitis

Underdiagnosis of drug-induced fever leads to extensive investigation and prolongation of hospitalization, and may lead to multiple unnecessary invasive procedures and a wrong diagnosis. Azathioprine is a widely used immunosuppressive drug. We report a case of a 53-year-old female patient diagnosed with autoimmune hepatitis treated with azathioprine, who presented to the emergency room with a 6-wk history of fever and chills without other associated symptoms. Since the patient's fever was of unknown origin, she was hospitalized. All treatment was stopped and an extensive workup to explore the source of fever and chills was performed. Results of chest X-ray, viral, urine, and blood cultures, autoimmune serology, transthoracic and transesophageal echocardiography, and abdominal ultrasound revealed no source of infection. A rechallenge test of azathioprine was performed and the fever and chills returned within a few hours. Azathioprine was established as the definite cause following rechallenge. Fever as an adverse drug reaction is often unrecognized. Azathioprine has been reported to cause drug-induced fever in patients with inflammatory bowel disease, rheumatoid arthritis, and sarcoidosis. To the best of our knowledge there have been no previous reports documenting azathioprine-induced fever in patients with autoimmune hepatitis. The occurrence of fever following the readministration of azathioprine suggests the diagnosis of drug-induced fever, particularly after the exclusion of other causes. A careful rechallenge is recommended to confirm the diagnosis.

The Utility of Tissue Immunohistochemistry and Viral Culture in Immunocompromised Patients Based on Endoscopic Findings

Purpose: Immunosuppressed patients presenting with gastrointestinal signs and symptoms are often assessed for opportunistic infections with endoscopic mucosal biopsy for tissue immunohistochemistry and viral cultures. However, the utility of this practice is unclear. Methods: Using laboratory and pathology databases we retrospectively identified all patients >18 years old undergoing tissue viral cultures or viral immunohistochemistry (IHC) between January 2005 and November 2011. The results of IHC staining, viral culture results, and endoscopic findings were independently reviewed by different physicians who were not aware of the other results. The primary end point was the diagnostic yield of IHC staining and viral cultures in patients with endoscopically normal and abnormal GI mucosa. Results: A total of 154 patients, age 18 to 80, who underwent a total of 209 procedures (102 upper and 107 lower GI endoscopy) were reviewed. 77% of patients had documented use of an immunosuppressive medication at the time of endoscopy. Endoscopic exam showed 62/209(30%) with ulcerations, 97/209(46%) with non-ulcer abnormal mucosa, and 50/209(24%) with normal mucosa. Of all the specimens, 14/209(7%) were positive for viral cultures and 10/209(5%) had positive IHC staining consistent with viral infection. Of the 14 specimens with positive viral cultures, the endoscopic findings included 12 with ulcers, 2 with erosions, and 2 normal findings. Also, 7/14 of these specimens had positive IHC and 6/14 patients had documented positive viral serology prior to endoscopy. The yield of viral cultures in association with endoscopic findings were 12/62(19%) for ulcers, 2/97(2%) for non-ulcer abnormal mucosa and 2/50 (4%) for normal mucosa. Thus, the yield of positive viral culture in non-ulcerated mucosa biopsies was 4/147 (3%), and 2 out of 4 patients had documented positive viral DNA PCR prior to endoscopy. The yield of positive IHC staining in association with endoscopic findings were 9/62(10%) for ulcers, 1/97(1%) for non-ulcer abnormal mucosa, and 0/50 (0%) for normal mucosa. Thus, the yield of positive IHC staining in non-ulcerated mucosa biopsy was 1/147 (0.7%). Although this patient did not have positive viral culture result, the patient had documented positive serology study, prior to endoscopy. Therefore, the additional yield for IHC was zero. Conclusion: Endoscopic biopsies for IHC and viral cultures often yielded positive results in immunocompromised patients with ulcerated mucosa. However, the yield in the setting of non-ulcerated mucosa is very low and not likely to alter patient management.

Uuse of a dashboard based on rapid viral tests results for predicting and managing clinical services

Viral culture results (VCT) correlates well with the intensity of clinical services but they are not available real time. Rapid viral tests results (RVT) are available on the same day and can be used for real time predictions. This study reports the use of RVT for such prediction for a period of 145 weeks in Hong Kong (HK). Two acute hospitals successfully use them to manage the winter Influenza surge of 2009.

Betanodavirus infection in golden pompano, Trachinotus blochii, fingerlings cultured in deep-sea cage culture facility in Langkawi, Malaysia

Golden pompano, Trachinotus blochii is a new marine fish candidate for aquaculture in Malaysia. The fingerlings of the fish are not yet produced locally but imported from neighboring countries and Taiwan. In the present study we report the first molecular detection of betanodavirus from diseased golden pompano fingerlings cultured in a deep-sea cage culture facility in Langkawi, Malaysia. The virus caused the fish fingerlings to exhibit abnormal swimming behavior, dark skin coloration and loss of appetite prior to mass mortality. Histopathological examination revealed acute cell vacuolation in both brain and retina tissues of the affected fingerlings. Results of viral culture, RT-PCR and DNA sequencing further confirmed the role of betanodavirus infection in the outbreak. Although phylogenetic analysis and mean nucleotide sequence of RNA2 genome of the betanodavirus showed high similarity to RGNNV betanodavirus isolated from Taiwan, origin of infection is difficult to establish since the RGNNV is also commonly reported in the Southeast Asian region and has the widest range of host marine fish species. Nevertheless, the finding of the present study requires a review of procedures in importing fish fingerlings for sustainable growth of the marine aquaculture industry in Malaysia.

Azathioprine-lnduced Fever in Sarcoidosis

To report a case of drug-induced fever associated with azathioprine treatment in a patient with sarcoidosis. A 52-year-old man with pulmonary sarcoidosis presented to the emergency department with a 1-day history of fever (temperature 39.9 °C), chills, nausea, and vomiting. One week earlier, azathioprine 50 mg/day had been started for worsening dyspnea. The patient was admitted and evaluated for acute infectious processes. All of his home medications (hydroxychloroquine, prednisone, fluticasone/salmeterol, lovastatin, pantoprazole, zolpidem, ibandronate, albuterol), except prednisone, were held. Results of chest X-ray, viral cultures, and urine and blood cultures revealed no source of infection. The patient's temperature returned to normal within 30 hours after discontinuation of azathioprine; rechallenge was not performed. Fever as an adverse drug reaction is often unrecognized, particularly in medically complex patients. Azathioprine has been reported to cause drug fever in patients with inflammatory bowel disease and in those with rheumatoid arthritis; to our knowledge, there have been no previous reports documenting azathioprine-induced fever in patients with sarcoidosis. The chronological course of febrile response and defervescence is highly suggestive of drug-induced fever. The rapid resolution of fever after discontinuation of azathioprine suggests that it was the likely source of fever in this patient. If azathioprine is increasingly prescribed in patients with sarcoidosis, fever as an adverse reaction may become more common.

Effects of Early Oseltamivir Therapy on Viral Shedding in 2009 Pandemic Influenza A (H1N1) Virus Infection

Pandemic influenza (H1N1) 2009 is susceptible to oseltamivir. There are few reports on its clinical and virologic response to oseltamivir. During the pandemic containment response in Singapore, all patients with positive polymerase chain reaction (PCR) results for pandemic influenza (H1N1) 2009 were hospitalized, given oseltamivir for 5 days, and discharged when daily PCR results for combined nasal and throat swab samples became negative. Six patients had concurrent positive viral culture and PCR results. The median age of the first 70 consecutive patients was 26 years (interquartile range, 21-38 years); 60% were men, and 29% had comorbidity. The mean time (+/-SD) from illness onset to hospital admission was 3+/-2 days. Influenza-like illness was noted in 63% of patients. Fever occurred in 91%, cough in 88%, sore throat in 66%, and rhinorrhea in 53% of patients. The mean duration (+/-SD) of viral shedding from illness onset was day 6+/-2 days. Viral shedding persisted beyond 7 days in 37% of patients. Clinical features and viral shedding were similar between those with and without comorbidity, except the former had more cough and lower oxygen saturation. Patients receiving oseltamivir on days 1 to 3 of illness had significantly shorter viral shedding duration, compared with those treated from day 4 onwards (P < .05). The mean durations (+/-SD) of positive PCR and viral culture results were 5+/-8 and 4+/-18 days, respectively, for 6 patients with concurrent positive viral culture and PCR results. Prolonged viral shedding was noted in young immunocompetent adults with mild pandemic influenza (H1N1) 2009 despite receipt of oseltamivir. When prescribed during the first 3 days of illness, oseltamivir shortened the duration of viral shedding.