Identification Of Virus Infection Research Articles

Microfluidic Chip-Assisted Upconversion Luminescence Biosensing Platform for Point-of-Care Virus Diagnostics.

Epidemics caused by multiple viruses continue to emerge, which have brought a terrible impact on human society. Identification of viral infections with high sensitivity and portability is of significant importance for the screening and management of diseases caused by viruses. Herein, a microfluidic chip (MFC)-assisted upconversion luminescence biosensing platform is designed and fabricated for point-of-care virus detection. Upconversion nanoparticles with excellent stability are successfully synthesized as luminescent agents for optical signal generation in the portable virus diagnostic platform. The relevant investigation results illustrate that the MFC-assisted virus diagnostic platform possesses outstanding performance such as good integration, high sensitivity (1.12 pg mL-1), ease of use, and portability. In addition, clinical sample test result verifies its more prominent virus diagnostic properties than commercially available rapid test strips. All of these thrilling capabilities imply that the designed portable virus diagnostic platform has great potential for future virus detection applications.

Update Advances on C-Reactive Protein in COVID-19 and Other Viral Infections

Severe coronavirus disease 2019 (COVID-19) can manifest as a viral-induced hyperinflammation with multiorgan dysfunction. It has been documented that severe COVID-19 is associated with higher levels of inflammatory mediators than a mild disease, and tracking these markers may allow early identification or even prediction of disease progression. It is well known that C-reactive protein (CRP) is the acute-phase protein and the active regulator of host innate immunity, which is highly predictive of the need for mechanical ventilation and may guide escalation of treatment of COVID-19-related uncontrolled inflammation. There are numerous causes of an elevated CRP, including acute and chronic responses, and these can be infectious or non-infectious in etiology. CRP are normally lacking in viral infections, while adaptive immunity appears to be essential for COVID-19 virus clearance, and the macrophage activation syndrome may explain the high serum CRP contents and contribute to the disease progression. Nevertheless, for the assessment of host inflammatory status and identification of viral infection in other pathologies, such as bacterial sepsis, the acute-phase proteins, including CRP and procalcitonin, can provide more important information for guiding clinical diagnosis and antibiotic therapy. This review is aimed to highlight the current and most recent studies with regard to the clinical significance of CRP in severe COVID-19 and other viral associated illnesses, including update advances on the implication of CRP and its form specifically on the pathogenesis of these diseases. The progressive understanding in these areas may be translated into promising measures to prevent severe outcomes and mitigate appropriate treatment modalities in critical COVID-19 and other viral infections.

Acute Kidney Injury Among Children Admitted With Viral Rhabdomyolysis.

Infectious etiologies cause a large portion of pediatric rhabdomyolysis. Among pediatric patients with rhabdomyolysis, it is unknown who will develop acute kidney injury (AKI). We sought to test the hypothesis that a viral etiology would be associated with less AKI in children admitted with rhabdomyolysis than a nonviral etiology. In this single-center retrospective cohort study, patients <21 years of age admitted with acute rhabdomyolysis from May 1, 2010, through December 31, 2018, were studied. The primary outcome was development of AKI, defined by using the Kidney Disease: Improving Global Outcomes guidelines. The primary predictor was identification of viral infection by laboratory testing or clinical diagnosis. Covariates included age, sex, race, insurance provider, presence of proteinuria and myoglobinuria, and initial creatinine kinase and serum urea nitrogen. Routine statistics and multivariable logistic modeling were performed via SAS 9.4 (SAS Institute, Inc, Cary, NC). In total, 319 pediatric patients with rhabdomyolysis were studied. The median age was 13 years. Patients were predominately male (69.9%), non-Hispanic Black (55.2%), and publicly insured (45.1%). We found no difference in the rates of AKI in those with a viral diagnosis versus those without a viral diagnosis (30 of 77 [39.0%] vs 111 of 234 [47.4%]; P = .19). Multivariable analysis revealed that viral diagnosis was not associated with the development of AKI. Patients ≥13 years of age, male patients, and those with proteinuria and elevated serum urea nitrogen on admission had increased odds of developing AKI. In our study, viral rhabdomyolysis did not have lower rates of AKI compared with nonviral etiologies of AKI; therefore, providers should consider continued caution in these patients.

Early Identification of COVID-19 Infection Using Remote Cardiorespiratory Monitoring: Three Case Reports.

BackgroundThe adoption of remote patient monitoring (RPM) in routine medical care requires increased understanding of the physiologic changes accompanying disease development and the proactive interventions that will improve outcomes.ObjectiveThe aim of this study is to present three case reports that highlight the capability of RPM to enable early identification of viral infection with COVID-19 in patients with chronic respiratory disease.MethodsPatients at a large pulmonary practice who were enrolled in a respiratory RPM program and who had contracted COVID-19 were identified. The RPM system (Spire Health) contains three components: (1) Health Tags (Spire Health), undergarment waistband-adhered physiologic monitors that include a respiratory rate sensor; (2) an app on a smartphone; and (3) a web dashboard for use by respiratory therapists. The physiologic data of 9 patients with COVID out of 1000 patients who were enrolled for monitoring were retrospectively reviewed, and 3 instances were identified where the RPM system had notified clinicians of physiologic deviation due to the viral infection.ResultsPhysiologic deviations from respective patient baselines occurred during infection onset and, although the infection manifested differently in each case, were identified by the RPM system. In the first case, the patient was symptomatic; in the second case, the patient was presymptomatic; and in the third case, the patient varied from asymptomatic to mildly symptomatic.ConclusionsRPM systems intended for long-term use and that use patient-specific baselines can highlight physiologic changes early in the course of acute disease, such as COVID-19 infection. These cases demonstrate opportunities for earlier diagnosis, treatment, and isolation. This study supports the need for further research into how RPM can be effectively integrated into clinical practice.

Identification of Virus Infection on Native Orchids in Liwa Botanical Garden

Liwa Botanical Garden is one of the regional botanical gardens in Indonesia with the theme “Indonesian Ornamental Plants”. At the present, disease infections are still a major obstacle in efforts to preserve and develop the potential natural orchids. Based on previous research, it is known that some individual orchids exhibit symptoms of viral disease infections, namely mosaic, chlorotic, streak, and necrosis. This research was conducted to determine the identification of against virus infections in Liwa Botanical Garden through a collection of samples that showed symptoms of infection, analysis of disease symptoms, and analysis of the level of disease resistance. The results showed the response of native orchids in Liwa Botanical Garden to viral infections showed symptoms in the form of necrotic black and chlorotic patches, yellowing leaves on orchids Coelogyne sp., Flickingeria sp., Calanthe sp., Trixspermum centipeda, Bulbophyllum sp. The type of orchid that shows the most symptoms is Flickingeria sp. a total of 8 samples. The type of natural orchid in the Liwa Botanical Garden which is most vulnerable to being infected with a virus is Flickingeria sp. with a disease intensity of 42%, while the type of orchid most infected virus is Coelogyne sp. with an intensity of 15%. The results of this activity are expected to be basic information in efforts to protect plants against diseases to support the application of conservation of natural orchids in the Liwa Botanical Garden.

Rt-Pcr In Diagnosis of Viral Infection

The main aim of this study is to assist in the diagnosis of viral infection using RT-PCR, which helps in easy detection and rapid result of infection. RT-PCR is more sensitive and precise to the identification of viral infection than the traditional process. It is a gold standard technique that helps to diagnose viral infection in the laboratory. Using different labeled samples ensures simple identification when used in multiple formats. RT-PCR viral nucleic acid testing was one of the fastest established methods of laboratory diagnosis. In a novel viral pandemic as COVID viral infection is an epidemic in the current situation. RT-PCR is helpful in the development dimension and the analysis aims to educate the scientist on the current state of viral infection. We explain both the benefits of using PCR and limitations. The PCR has provided major methodological advantages and improved patient outcomes. This might help in the detection of viral infection at a faster rate.

Simple, fast and affordable triaging pathway for COVID-19

Coronavirus disease 2019 has caused a global pandemic. The majority of patients will experience mild disease, but others will develop a severe respiratory infection that requires hospitalisation. This is causing...

DsRNA-Seq: Identification of Viral Infection by Purifying and Sequencing dsRNA.

RNA viruses are a major source of emerging and re-emerging infectious diseases around the world. We developed a method to identify RNA viruses that is based on the fact that RNA viruses produce double-stranded RNA (dsRNA) while replicating. Purifying and sequencing dsRNA from the total RNA isolated from infected tissue allowed us to recover dsRNA virus sequences and replicated sequences from single-stranded RNA (ssRNA) viruses. We refer to this approach as dsRNA-Seq. By assembling dsRNA sequences into contigs we identified full length or partial RNA viral genomes of varying genome types infecting mammalian culture samples, identified a known viral disease agent in laboratory infected mice, and successfully detected naturally occurring RNA viral infections in reptiles. Here, we show that dsRNA-Seq is a preferable method for identifying viruses in organisms that don’t have sequenced genomes and/or commercially available rRNA depletion reagents. In addition, a significant advantage of this method is the ability to identify replicated viral sequences of ssRNA viruses, which is useful for distinguishing infectious viral agents from potential noninfectious viral particles or contaminants.

Acute phase reactant serum amyloid A in inflammation and other diseases.

Acute-phase reactant serum amyloid A (A-SAA) plays an important role in acute and chronic inflammation and is used in clinical laboratories as an indicator of inflammation. Although both A-SAA and C-reactive protein (CRP) are acute-phase proteins, the detection of A-SAA is more conclusive than the detection of CRP in patients with viral infections, severe acute pancreatitis, and rejection reactions to kidney transplants. A-SAA has greater clinical diagnostic value in patients who are immunosuppressed, patients with cystic fibrosis who are treated with corticoids, and preterm infants with late-onset sepsis. Nevertheless, for the assessment of the inflammation status and identification of viral infection in other pathologies, such as bacterial infections, the combinatorial use of A-SAA and other acute-phase proteins (APPs), such as CRP and procalcitonin (PCT), can provide more information and sensitivity than the use of any of these proteins alone, and the information generated is important in guiding antibiotic therapy. In addition, A-SAA-associated diseases and the diagnostic value of A-SAA are discussed. However, the relationship between different A-SAA isotypes and their human diseases are mostly derived from research laboratories with limited clinical samples. Thus, further clinical evaluations are necessary to confirm the clinical significance of each A-SAA isotype. Furthermore, the currently available A-SAA assays are based on polyclonal antibodies, which lack isotype specificity and are associated with many inflammatory diseases. Therefore, these assays are usually used in combination with other biomarkers in the clinic.

Polyethylene Glycol Accelerates Loop-mediated Isothermal Amplification (LAMP) Reaction

Loop-mediated isothermal amplification (LAMP) has several advantages: this technique involves gene amplification under isothermal conditions using only one high-specificity enzyme; the amplification efficiency is so high that large quantities of pyrophosphoric acid are formed as a by-product of DNA synthesis; furthermore, the results can be judged directly on the basis of turbidity. On the other hand, a PCR requires approximately 3.5-4.0 hours. The LAMP method is faster than the PCR method and is also relatively inexpensive. In the present study, we modified the composition of the reaction solutions to reduce the LAMP reaction time; more specifically, a thickener, either polyethylene glycol 8000 or 20000, was added. These results showed that the LAMP method was faster than the original method, and it is able to detect both turbidity and fluorescence. In conclusion, the LAMP reaction could be performed within 20 minutes when reaction mixture supplemented with a thickener was used. This method can be used for tests in various fields such as the diagnosis of hereditary disease and identification of viral infections as point-of-care testing.

Serum protein profiles as potential biomarkers for infectious disease status in pigs

BackgroundIn veterinary medicine and animal husbandry, there is a need for tools allowing the early warning of diseases. Preferably, tests should be available that warn farmers and veterinarians during the incubation periods of disease and before the onset of clinical signs. The objective of this study was to explore the potential of serum protein profiles as an early biomarker for infectious disease status. Serum samples were obtained from an experimental pig model for porcine circovirus-associated disease (PCVAD), consisting of Porcine Circovirus type 2 (PCV2) infection in combination with either Porcine Parvovirus (PPV) or Porcine Reproductive and Respiratory Syndrome virus (PRRSV). Sera were collected before and after onset of clinical signs at day 0, 5 and 19 post infection. Serum protein profiles were evaluated against sera from non-infected control animals.ResultsProtein profiles were generated by SELDI-TOF mass spectrometry in combination with the Proteominer™ technology to enrich for low-abundance proteins. Based on these protein profiles, the experimentally infected pigs could be classified according to their infectious disease status. Before the onset of clinical signs 88% of the infected animals could be classified correctly, after the onset of clinical sigs 93%. The sensitivity of the classification appeared to be high. The protein profiles could distinguish between separate infection models, although specificity was moderate to low. Classification of PCV2/PRRSV infected animals was superior compared to PCV2/PPV infected animals. Limiting the number of proteins in the profiles (ranging from 568 to 10) had only minor effects on the classification performance.ConclusionsThis study shows that serum protein profiles have potential for detection and identification of viral infections in pigs before clinical signs of the disease become visible.

Viral Infections in the Neonate

Maternal to child transmission of viral infections can have devastating outcomes on the developing fetus and the neonate. Effective prevention and screening programs for women of childbearing age are key factors in influencing the outcomes for the neonate. Furthermore, early identification of viral infections in neonates is critical to the success of the treatment plan. This article describes the mechanism of maternal to child transmission of viral infections and examines the general approach to the neonate with suspected viral infection. The diagnosis, treatment, and prevention of neonatal cytomegalovirus, herpes simplex virus, and human immunodeficiency virus infections are presented. Lastly, current challenges and controversies related to the management of viral infections in the neonate are discussed.

Identification of viral infections in the prostate and evaluation of their association with cancer

BackgroundSeveral viruses with known oncogenic potential infect prostate tissue, among these are the polyomaviruses BKV, JCV, and SV40; human papillomaviruses (HPVs), and human cytomegalovirus (HCMV) infections. Recently, the Xenotropic Murine Leukemia Virus-related gammaretrovirus (XMRV) was identified in prostate tissue with a high prevalence observed in prostate cancer (PC) patients homozygous for the glutamine variant of the RNASEL protein (462Q/Q). Association studies with the R462Q allele and non-XMRV viruses have not been reported. We assessed associations between prostate cancer, prostate viral infections, and the RNASEL 462Q allele in Mexican cancer patients and controls.Methods130 subjects (55 prostate cancer cases and 75 controls) were enrolled in the study. DNA and RNA isolated from prostate tissues were screened for the presence of viral genomes. Genotyping of the RNASEL R462Q variant was performed by Taqman method.ResultsR/R, R/Q, and Q/Q frequencies for R462Q were 0.62, 0.38, and 0.0 for PC cases and 0.69, 0.24, and 0.07 for controls, respectively. HPV sequences were detected in 11 (20.0%) cases and 4 (5.3%) controls. XMRV and HCMV infections were detected in one and six control samples, respectively. The risk of PC was significantly increased (Odds Ratio = 3.98; 95% CI: 1.17-13.56, p = 0.027) by infection of the prostatic tissue with HPV. BKV, JCV, and SV40 sequences were not detected in any of the tissue samples examined.ConclusionsWe report a positive association between PC and HPV infection. The 462Q/Q RNASEL genotype was not represented in our PC cases; thus, its interaction with prostate viral infections and cancer could not be evaluated.

DNA polymerase gene locus of Cercopithecine herpesvirus 1 is a suitable target for specific and rapid identification of viral infection by PCR technology

The family Herpesviridae comprises at least 100 herpesviruses. Numerous human and animal pathogenic herpesviruses have been identified so far, including Cercopithecine herpesvirus 1 (CeHV-1). This virus is a member of the subfamily Alphaherpesvirinae and is the most hazardous herpesvirus to man. CeHV-1 is also known as B-virus or monkey B virus and as Herpesvirus simiae. In order to gain more genetic information, the viral DNA polymerase (DPOL) gene was identified using polymerase chain reaction (PCR) and DNA nucleotide sequence analysis. The deduced amino acid sequence contains the motifs and signatures that are typical for the B-family of DPOLs. The DPOL gene of CeHV-1 was found to be a suitable target for the specific and rapid identification of the Cercopithecine herpesvirus 1 infection by PCR technology. Comparative analysis of the DNA sequences of the DPOL gene loci of CeHV-1, Human herpesvirus 1 and 2 (HHV-1 and HHV-2), and other herpesviruses was carried out for determination of unique genomic regions of the individual DPOL genes. A primer set of 12 primers was used for screening the DNA of CeHV-1, HHV-1, and HHV-2 by detailed PCR. It was found that six out of twelve primer combinations are able to detect specifically the CeHV-1 genome without cross reactivity with the genome of HHV-1 and/or HHV-2. The specificity of the individual amplified DNA fragments was confirmed by DNA nucleotide sequence analysis. The results of these studies indicate that the six primer combinations of the specific CeHV-1 DPOL primer set is the method of choice for a rapid, precise and specific identification of a CeHV-1 infection by PCR. Due to the fact that this specific CeHV-1 DPOL primer set does not amplify any DNAs of HHV-1 or HHV-2 genome this technology is stressing and can be successfully used unlimited and more credible in all laboratories with PCR technical facility routinely for detection of a CeHV-1 infection in vivo or in vitro.

Six viruses identified in potato plants grown in Alabama, U.S.A.

A survey was conducted for virus-infected potato plants in commercial production fields in Baldwin, Cullman, DeKalb, and Jackson counties in Alabama. Infected plants were assessed visually, i.e., on the basis of occurrence of virus-like symptoms, and by enzyme-linked immunosorbent assay (ELISA). In 1997, potato plants were tested for Cucumber mosaic virus (CMV), Potato virus Y (PVY) and Tobacco etch virus (TEV). PVY was detected from foliar samples collected from Baldwin, Cullman, and Jackson counties. Neither CMV nor TEV were detected in any of the samples. In 1998, potato plants were once again tested for the presence of CMV, PVY, and TEV, with additional analyses for Potato viruse A (PVA), Potato M (PVM), Potato S (PVS), Potato X (PVX), and Potato leafroll virus (PLRV). Four viruses were detected from potato leaf samples collected from Baldwin County: PVS, PVX, PVY, and PLRV. Samples collected from DeKalb and Jackson counties were shown to be infected with CMV, PVA, PVS, PVX, and PVY. Identification of virus infection based on visual assessment was relatively inconsistent with the exception of PLRV-infected potato plants, which had upward curled leaves. The expression of infection by PVA, PVS, PVX, and PVY varied; some infected potato plants were symptomless while others had reddening of leaves that sometimes included a mild mosaic. This is the first report to identify viruses in commercially-grown potato plants in Alabama.

32] Identification of viral infection by confocal microscopy

Confocal microscopy is a valuable adjunct to electron microscopy in the fields of diagnostic and investigative virology. Confocal imaging can be used to examine large amounts of tissue stained by a variety of methods for evidence of viral infection. Areas thus identified can then be processed for ultrastructural study, allowing a highly focused search for viral pathogens. With the possible exception of the vibrating microtome, all of the equipment and reagents necessary for the preparation of specimens for confocal scanning are available in any well-stocked histology laboratory. Although originally developed to facilitate viral diagnosis by EM, the methods described herein can be applied to the ultrastructural study of any focal pathologic process.

Detection of Epstein-Barr virus DNA in virus-infected cells by electron microscopic in situ hybridization.

We describe a procedure for in situ hybridization using a biotinylated Epstein-Barr virus (EBV) sequence with detection at the light and electron microscopic levels. In situ hybridization using an immunogold-silver staining detection system was used to identify biotinylated DNA probes in cell smears and in Lowicryl K4M-embedded EBV-infected and -noninfected cell lines. At the light microscopic level, the reaction product of hybridized EBV DNA sequence seemed to be located mainly in the nuclei. The labelling was dependent on the cell strains. However, at the electron microscopic level, the reaction product was evident as spots or clusters distributed not only in the nuclei of EBV-infected cells but also in the cytoplasm and extracellular particles. These findings suggest that immature particles in the cytoplasm contain EBV DNA. This procedure can be applied to the observation and identification of virus infection.

Viral pathogens and clinical manifestations associated with acute lower respiratory tract infections in children of the Sudan

Background: Incomplete knowledge regarding the viral agents causing respiratory infections in children living in developing countries impedes diagnosis and management of patients. Objectives: To assess the role of viral pathogens in Sudanese children presenting with acute lower respiratory tract infections (ALRI). Study design: The study population consisted of patients presenting with ALRI at the Children's Emergency Hospital in Khartoum during 2 periods (December 1987 to April 1988 and September 1990 to March 1991). Identification of viral infections was based an antigen detection by immunofluorescence and enzyme-linked immunosorbent assay (ELISA) on nasopharyngeal secretions and/or serology. Results: After exclusion of children with measles, 102 and 111 children, respectively, were prospectively enrolled in the study during the 2 periods. Their ages ranged between one mouth and 14 years (mean 2.0 years). Radiologic pulmonary infiltrations were detected in 135 (66%) of the 206 patients who had chest radiographs, whereas 7 (3%) showed lobar pneumonia. The case fatality rate was 2.3%. Of 83 virus infections detected, 79 were in children < years and consisted mainly of respiratory syncytial virus (RSV, 28%), followed by parainfluenza (7%), adenovirus (5%) and influenza A (2%). Infiltrates on radiographs were significantly less often found in virus-infected cases than in ALRI-cases with negative virus tests. Conclusions: RSV predominantly infected young infants (≤ 3 months) compared with those who were 5 years or older (28% and 13%, respectively). A tendency for respiratory viruses to be more prevalent was seen during the cooler months (January and February). There were no significant differences in clinical features between virus-positive cases and others. The panorama of viral infections proved to be the sme as in other developing countries.

Fungal transmission of a potyvirus: uredospores of Puccinia sorghi transmit maize dwarf mosaic virus.

Maize dwarf mosaic virus (MDMV) and maize rust, Puccinia sorghi Schw., occur as natural infections on cultivated maize in South Africa. P. sorghi often occurs as a secondary late infection on maize plants which have already been infected with MDMV earlier in the season, either seed or aphid transmitted. When MDMV isolates from maize plants naturally infected by both virus and fungus were propagated by sap inoculation in plant growth rooms, residual uredospores in the sap gave rise to the development of uredia under conditions of high humidity. When uredospores developing on MDMV-B-infected plants were germinated on virus free maize seedlings, these plants became infected with MDMV-B. Similarly, when uredospores, originating from maize plants infected with MDMV-A, were scattered onto virus free maize seedlings, these plants became infected with MDMV-A. The presence of virus on uredospores in infected plant tissue was visualized by indirect immunofluorescence. Identification of virus infection was by DAS-ELISA and immunoelectro-blotting utilizing strain-specific antisera. Virus transmission occurred between closely situated plants which had no actual contact (unaided transmission). MDMV-B transmission by uredospores, to new maize seedlings, has been maintained for three successive years (1988-1991) in a plant growth room. The MDMV-B isolate remained sap and non-persistently aphid transmissible.

Ultrastrctural identification of viral infections in paraffin sections stained by immunohistochemistry and in situ hybridization

Confirmation of viral infections in surgical biopsies is sometimes limited to routine paraffin sections because a specimen may not have been set aside for culture or culture results are negative. Since viral infections are frequently focal and localized to only a few cells, definitive histopathological diagnosis can be difficult to achieve even with immunohistoGhemistry (IHC), in situ hybridization (ISH) or electron microscopy. In this paper, we would like to demonstrate a siirple and direct approach to confirming viral infections in paraffin sections , using two immuno-labelling techniques coupled with electron microscopy.Paraffin sections of surgical specimens are deparaffinized in xylene and rehydrated through graded ethyl alcohol. Herpes simplex virus (Figure 1) and Toxoplasma gondii are localized immunohistochemically by the peroxidase antiperoxidase technique and Pneumocystis carinii (Figure 2) by 3 step ipdirect method, using 3,3'-diaminobenzidine tetrahydrochloride as the chrmogen, CMV ENA probe (ENZO Biochemicals) is used for in situ hybridization, following kit instructions (Figure 3 &amp; 4). After the final chromogen reaction, wet sections are examined with a light microscope. Slides in vrfiich focal positive staining is difficult to interpret as specific are fixed in osmium tetroxide, dehydrated with graded acetone, and infiltrated with Poly/Bed 812. After polymerization, the epoxy blocks containing area of interest can be easily separated from the glass slides by heating on a 100 C hotplate for approximately 20 seconds . The area of interest should be positioned in the center of the block face for ultrathin sectioning. With careful adjustments of knife and microtcme, the area of interest can be precisely sectioned without loss of tissue. Ultrathin sections are collected on uncoated mesh grids for higher contrast and then directly examined by TEM. Peroxidase-labelled microorganisms or viral DHA can be easily localized and identified without heavy metal staining. Cellular details can be further increased by staining with uranyl acetate and lead citrate. Tissue remaining in block face after sectioning is available for IM comparison with the EM micrographs.