Detection Of Viral Antigens Research Articles

Ultrasound-guided minimally invasive autopsy as a tool for rapid post-mortem diagnosis in the 2018 Sao Paulo yellow fever epidemic: Correlation with conventional autopsy.

BackgroundNew strategies for collecting post-mortem tissue are necessary, particularly in areas with emerging infections. Minimally invasive autopsy (MIA) has been proposed as an alternative to conventional autopsy (CA), with promising results. Previous studies using MIA addressed the cause of death in adults and children in developing countries. However, none of these studies was conducted in areas with an undergoing infectious disease epidemic. We have recently experienced an epidemic of yellow fever (YF) in Brazil. Aiming to provide new information on low-cost post-mortem techniques that could be applied in regions at risk for infectious outbreaks, we tested the efficacy of ultrasound-guided MIA (MIA-US) in the diagnosis of patients who died during the epidemic.Methodology/principal findingsIn this observational study, we performed MIA-US in 20 patients with suspected or confirmed YF and compared the results with those obtained in subsequent CAs. Ultrasound-guided biopsies were used for tissue sampling of liver, kidneys, lungs, spleen, and heart. Liver samples from MIA-US and CA were submitted for RT-PCR and immunohistochemistry for detection of YF virus antigen. Of the 20 patients, 17 had YF diagnosis confirmed after autopsy by histopathological and molecular analysis. There was 100% agreement between MIA-US and CA in determining the cause of death (panlobular hepatitis with hepatic failure) and main disease (yellow fever). Further, MIA-US obtained samples with good quality for molecular studies and for the assessment of the systemic involvement of the disease. Main extrahepatic findings were pulmonary hemorrhage, pneumonia, acute tubular necrosis, and glomerulonephritis. One patient was a 24-year-old, 27-week pregnant woman; MIA-US assessed the placenta and provided adequate placental tissue for analysis.ConclusionsMIA-US is a reliable tool for rapid post-mortem diagnosis of yellow fever and can be used as an alternative to conventional autopsy in regions at risk for hemorrhagic fever outbreaks with limited resources to perform complete diagnostic autopsy.

Detection of Antigens of Cardiotropic Viruses in Atherosclerotic Plaques in Patients with Fatal Myocardial Infarction

To assess the frequency of detection of cardiotropic virus antigens in coronary artery atherosclerotic plaques in patients with fatal myocardial infarction (MI). We examined fragments of coronary plaques of 12 patients with fatal type 1 MI. Immunohistochemistry (IHC) of plaques was performed with the paraffin blocks using antibodies to Herpes simplex virus (HSV)-1, HSV-2, HSV-6, cytomegalovirus (CMV), parvovirus B19, adenovirus, Epstein-Barr virus and enteroviruses. According to the IHC all patients had virus antigens. The most common virus agents in fragments of coronary plaques were HSV-6 (10 patients) and enteroviruses (5 patients). Antigens of CMV, parvovirus B19, adenovirus, Epstein-Barr virus were not detected in any case. In this study viral antigens in coronary artery atherosclerotic plaques were found in all victims of fatal MI. There was no difference in the frequency of detection and type of viral agents between plaques in culprit arteries and uncomplicated atherosclerotic plaques.

Ultrasensitive Ebola Virus Antigen Sensing via 3D Nanoantenna Arrays.

Sensitive detection of pathogens is crucial for early disease diagnosis and quarantine, which is of tremendous need in controlling severe and fatal illness epidemics such as of Ebola virus (EBOV) disease. Serology assays can detect EBOV-specific antigens and antibodies cost-effectively without sophisticated equipment; however, they are less sensitive than reverse transcriptase polymerase chain reaction (RT-PCR) tests. Herein, a 3D plasmonic nanoantenna assay sensor is developed as an on-chip immunoassay platform for ultrasensitive detection of Ebola virus (EBOV) antigens. The EBOV sensor exhibits substantial fluorescence intensity enhancement in immunoassays compared to flat gold substrate. The nanoantenna-based biosensor successfully detects EBOV soluble glycoprotein (sGP) in human plasma down to 220 fg mL-1 , a significant 240 000-fold sensitivity improvement compared to the 53 ng mL-1 EBOV antigen detection limit of the existing rapid EBOV immunoassay. In a mock clinical trial, the sensor detects sGP-spiked human plasma samples at two times the limit of detection with 95.8% sensitivity. The results combined highlight the nanosensor's extraordinary capability of detecting EBOV antigen at ultralow concentration compared to existing immunoassay methods. It is a promising next-generation bioassay platform for early-stage disease diagnosis and pathogen detection for both public health and national security applications.

Evaluation of Animal Rabies Surveillance System, Ekiti State, Nigeria, 2012-2017

Evaluation of Animal Rabies Surveillance System, Ekiti State, Nigeria, 2012-2017

Purification of IgG against ribonucleoprotein by a homemade immunoaffinity chromatography column for rabies diagnosis

Polyclonal or monoclonal antibodies against rabies virus ribonucleoprotein (RNP) conjugated to fluorescein isothiocyanate (FITC) have been employed for Rabies virus (RABV) antigen detection by the direct fluorescent antibody test (DFA). To date, these biomolecules have been purified by traditional methods such as precipitation by ammonium sulfate or ion exchange chromatography followed by ammonium sulfate precipitation, which allows only for partial detection of the protein of interest. In this study, we aimed to purify anti-RNP polyclonal horse IgG antibodies by cation-exchange chromatography in combination with a homemade immunoaffinity chromatography on RNP immobilized (RNP-IAC). Furthermore, to evaluate the accuracy of the prepared anti-RNP IgG fluorescent antibody in diagnostic purposes, DFA was applied for RABV antigen detection in suspected brain samples of different animal species. The combination of these two techniques made it possible to obtain antibodies with high selectivity and purity. Compared with the performance of the traditional method, anti-RNP IgG antibodies purified by RNP-IAC can be obtained from a smaller volume of hyperimmune serum and with greater avidity. Furthermore, the results obtained by DFA analyses revealed that the prepared anti-RNP IgG fluorescent antibody achieved 100% diagnostic specificity and sensitivity for RABV antigen detection. Thus, two-technique chromatographic, including RNP-IAC technology could be appropriate methods for the purification of polyclonal anti-RNP IgG for the use as a diagnostic reagent for rabies.

Optimization and evaluation of a non-invasive tool for peste des petits ruminants surveillance and control

Peste des petits ruminants (PPR) is a highly contagious and devastating viral disease affecting mainly sheep and goats, but also a large number of wild species within the order Artiodactyla. A better understanding of PPR transmission dynamics in multi-host systems is necessary to efficiently control the disease, in particular where wildlife and livestock co-occur. Notably, the role of wildlife in PPR epidemiology is still not clearly understood. Non-invasive strategies to detect PPR infection without the need for animal handling could greatly facilitate research on PPR epidemiology and management of the disease in atypical hosts and in complex field situations. Here, we describe optimized methods for the direct detection of PPR virus genetic material and antigen in fecal samples. We use these methods to determine the detection window of PPR in fecal samples, and compare the sensitivity of these methods to standard invasive sampling and PPR diagnostic methods using field samples collected at a wildlife-livestock interface in Africa. Our results show that quantitative reverse transcription PCR (RT-QPCR) amplification of PPRV from fecal swabs has good sensitivity in comparison to ocular swabs. Animals infected by PPRV could be identified relatively early on and during the whole course of infection based on fecal samples using RT-QPCR. Partial gene sequences could also be retrieved in some cases, from both fecal and ocular samples, providing important information about virus origin and relatedness to other PPRV strains. Non-invasive strategies for PPRV surveillance could provide important data to fill major gaps in our knowledge of the multi-host PPR epidemiology.

Development of a multiplexed antigen detection immunoassay for detection of viral agents

Purpose: The spread of infectious disease continues to present a challenge for modern global public health initiatives, as was evidenced by the recent Ebola outbreak in West Africa. In disease outbreaks, diagnostics are the first line of defense in identifying the causative agent, treatment decisions, and control and prevention of future outbreaks. Methods & Materials: We designed a multiplexed immunoassay centered on detection of viral antigens in serum/plasma samples for EBOV, LASV, MARV, CCHFV, RVFV, alphaviruses, and flaviviruses. This flexible, immunoassay system, based on the MAGPIX® platform has faster sample-to-answer time over traditional methods and higher throughput, as assays can be multiplexed. Assays were developed and validated against live and inactivated viral supernatants and other antigen types. Inclusivity and exclusivity panels were run to ensure no crossreactivity between assays. Results: Each assay was able to detect the appropriate target, whether in inactivated, recombinant, and live agent form. Limits of detection were generated in plaque forming units (pfu) per ml of live agent. Limits of detection were also established for appropriate recombinant reagents when available, as these reagents serve as the positive controls for the multiplexed assays when deployed to overseas partners. Conclusion: Development of robust and sustainable immunoassays is important as they have great potential for use during outbreak events to provide a more complete picture of the presence of viral agents. Immunoassays are capable of providing further insight to shape outbreak response. The recent Ebola outbreak demonstrated the great sensitivity of RT-PCR, but often patients remained positive by PCR well after other symptoms subsided. An integrated diagnostic system that uses sensitive and specific RT-PCR in combination with higher throughput, multiplexed immunodiagnostic methods can provide the highest confidence in a diagnostic result.

Laser scanning optical investigations of dengue virus infection from antibody fluorescence of human blood

Dengue is the most important arthropod-borne viral disease of public health significance today, and the geographic distribution includes more than 100 countries worldwide. A definite diagnosis of dengue virus infection is based on clinical criteria supported by antibody tests of whole blood. In this study, we report on dengue infected whole blood diagnostics using laser scanning confocal microscopy (LSCM) over a laser excitation of 488, 543 and 633 nm wavelengths. Based on our findings, the system has potential applications in the detection and quantification of dengue virus infected cells, antigens and antibodies in blood in vitro. Analysis of about 100 infected patients shows that the primary organism (or pathogen), characteristic of dengue infection, causes a significant biochemical change in the nature of human blood in which it resides, and, due to which, fluorescence increases in dengue samples. Thus, the use of a reliable and sensitive rapid diagnostic test using LSCM can simultaneously detect the presence of anti-dengue IgG and IgM antibodies, which is a breakthrough in diagnosis and of great clinical use. The antibodies for dengue continue to be distinguishable for a couple of months after infection. This change in blood fluorescence may be a significant contribution from light scattering through antibodies produced against the dengue antigen.

The relationship between rhinovirus infection and acute wheezing in young children with recurrent wheezing

Asthma is a common chronic respiratory disease in childhood. The present study aims to assess the association between rhinovirus (RV) infection and acute wheezing in the occurrence of recurrent wheezing in 5-year old and younger children. A total of 109 children with recurrent wheezing and 70 children without wheezing (controls) were recruited from October 2013 to March 2015. Nasopharyngeal aspirate samples were obtained from all children. RV, human metapneumovirus (hMPV), and bocavirus (BoV) were tested by reverse transcription-polymerase chain reaction.Respiratory syncytial virus (RSV), parainfluenza virus (PIV), influenza virus (IV), and adenoviruses (ADV) were confirmed by detection of viral antigens via fluoroimmunoassay. Viral infection was more commonly detected in children with recurrent wheezing than in controls (odds ratio (OR): 6.10; 95% confidence interval (CI): 2.89–12.87). RV and RSV were found in both wheezing children and controls. However, RV was detected more in wheezing children than in controls (OR: 3.07; 95% CI: 1.37–6.90), followed by RSV (OR: 5.33; 95% CI: 1.53–18.62). Furthermore, RV more tended to infect wheezing children after infancy. Compared with children infected with other viruses, higher levels of eosinophil were found in wheezing children with RV infection ( P < 0.05). RV was the main pathogen that induced exacerbation in young children with recurrent wheezing. Furthermore, the rate of RV infection was higher in children above 1 year old. RV infection might be associated with high levels of eosinophil.

Standardization and application of immunofluorescence and immuno- peroxidase tests for detection of bluetongue virus antigen

India is enzootic for bluetongue (BT), a predominant disease of small ruminants. The most important task in the control of disease is rapid and sensitive detection of virus. The present study was undertaken to standardize immunofluorescence (IFT) and immunoperoxidase tests (IPT) employing BTV serogroup specific VP7 monoclonal antibodies (MAbs), polyclonal homologous, and polyclonal heterologous antisera against specific serotypes of BTV for detection of BTV antigen. Serial tenfold dilutions of BTV-9 were tested for limit of detection (LoD) of IFT, IPT, and molecular assays by using MAbs against VP7, homologous anti-BTV-9 serum, and heterologous anti-BTV-16 serum. LoD of IFT was found to be 101 TCID50/mL using MAbs against VP7, anti BTV-9 serum, and anti BTV-16 serum. LoD of IPT was found to be 101 TCID50/mL, 102 TCID50/mL, and 102 TCID50/mL using MAbs against VP7, anti BTV-9 serum and anti BTV-16 serum, respectively. LoD of RT-PCR was 101 TCID50/mL and that of real time PCR was 100 TCID50/mL. This standardized assay was then applied for BTV detection in BTV suspected field samples collected from BT outbreaks followed by confirmation with virus isolation and NS3 group specific PCR. The current study shows that IFT and IPT are specific tests for group specific BTV identification. For IFT, monoclonal and polyclonal (homologous and heterologous) source of antibodies had similar sensitivity in the ability of BTV detection; whereas the most sensitive mode of detection by IPT was with MAbs.

NFKB1 promoter polymorphism: A new predictive marker of cytomegalovirus infection after kidney transplantation.

Cytomegalovirus (CMV) infection represents a common cause of morbidity and mortality in kidney transplant recipients (KTR). The NF-kB signaling pathway is highly involved in the pathogenesis of CMV infection. The -94ins/delATTG functional polymorphism in the promoter of NFKB1 has been associated with low intracellular levels of the protein and high incidence of inflammatory and autoimmune disease. In this study, we evaluated the association of this NFKB1 polymorphism with the risk of CMV infection. CMV infection was defined as virus isolation or detection of viral antigens or nucleic acid in any body fluid or tissue specimen. Using Cox regression and survival analysis, we analyzed the association between the polymorphism and CMV infection as well as recurrence in the first 12months after transplantation. We analyzed the -94ins/delATTG NFKB1 polymorphism of 189 KTRs. The 65% of CMV infections occurred in ins/ins group. Survival free from CMV infection was 54.7% for ins/ins group and 79.4% for deletion carriers one year after transplantation (P<0.0001). At multivariate regression, deletion carriers showed a lower risk of CMV infection and recurrence with respect to ins/ins KTRs (HR=0.224 P=0.0002; HR=0.307, P=0.012, respectively). In conclusion, pretransplantation screening for NFKB1 -94ins/delATTG polymorphism may predict CMV infection and improve the management of patients at higher risk of infection in the post-transplant period.

A STUDY OF DEMOGRAPHIC PROFILE ASSOCIATION WITH SEROPOSITIVE CASES OF DENGUE VIRUS INFECTION IN LAHORE

Introduction: Dengue fever is caused by a flavi virus, called dengue virus. Aedes Egypti and Aedes Albopictus transmit this virus. Diagnosis is through direct i.e. virus isolation, genome detection and antigen detection and indirect i.e. serology IgG and IgM detection methods.&#x0D; Objective: The objective of the study was to study the association of different epidemiological factors with dengue fever in Lahore.&#x0D; Material and Methods: 300 suspected dengue cases presenting in Emergency, Dengue isolation ward, different wards and OPD’s of Mayo Hospital, Sir Ganga Ram Hospital and Jinnah Hospital Lahore were obtained.&#x0D; Sampling: Convenience purposive sampling was opted.&#x0D; Study Design: It was analytical cross sectional. The different demographic and epidemiological factors were noted to see their association with dengue fever.&#x0D; Results: 90 patients (30% of total) were positive for IgM and 210(70 % of total) were negative for IgM. 108 patients (36% of total)were positive for IgG and 192(64% of total) were negative for IgG . 54 (18%) were positive for IgM and 63 (21%) males were positive for IgG. 36 females (12%) were positive for IgM and 45 (15%) females were positive for IgG. Simultaneous positivity of both IgG and IgM was in 18 male patients (6%) and in 21 female patients (7%). IgM positivity was more in age group 11-20 (9%) and least in age group 71-80. IgG positivity was more in age group 11-20 (12%) and least in age groups 0-11 and 71-80(2% each). Their mean age were 29.14 + 17.03 (SD) years ranging from 9 months to 90 years? Maximum positivity of IgM and IgG both was observed in Northern Lahore i.e. 48 (19 %). Minimum positivity of both IgM and IgG was observed in Eastern Lahore i.e. 3 (3 %) respectively.

Alston Virus, a Novel Paramyxovirus Isolated from Bats Causes Upper Respiratory Tract Infection in Experimentally Challenged Ferrets.

Multiple viruses with zoonotic potential have been isolated from bats globally. Here we describe the isolation and characterization of a novel paramyxovirus, Alston virus (AlsPV), isolated from urine collected from an Australian pteropid bat colony in Alstonville, New South Wales. Characterization of AlsPV by whole-genome sequencing and analyzing antigenic relatedness revealed it is a rubulavirus that is closely related to parainfluenza virus 5 (PIV5). Intranasal exposure of mice to AlsPV resulted in no clinical signs of disease, although viral RNA was detected in the olfactory bulbs of two mice at 21 days post exposure. Oronasal challenge of ferrets resulted in subclinical upper respiratory tract infection, viral shedding in respiratory secretions, and detection of viral antigen in the olfactory bulb of the brain. These results imply that AlsPV may be similar to PIV5 in its ability to infect multiple mammalian host species. This isolation of a novel paramyxovirus with the potential to transmit from bats to other mammalian species reinforces the importance of continued surveillance of bats as a source of emerging viruses.

Phylogenetic Analysis of MERSCoV in Human and Camels in Iraq

The present study was conducted to evaluate the genetic relationship among Middle East respiratory syndrome coronavirus (MERSCoV) of human and camels origin at the period from October 2015 to February 2016.One hundred samples were collected from camel and 100 from human. Nighty four from nasal swabs and six from oropharyngeal swabs Camel samples secerned by immunochromatographic assay (ICA) for detection of viral antigen. The total percentage of ICA positivity was 28%. Human and camel samples subjected to Revers transcription real time- PCR and carried out by RNA extraction by using specific primers and Taq- Man-Probe for detection of nucleocapsid gene 113 bp. The total positive result in camels were 15% ,there was no significant difference between sex and type of samples, in relation to the age group the results showed that age group more than ten years of camel was the heights percent. With significant difference at P&lt;0.05. According to the months of the year October recorded the highest infection rate with significant difference at p&lt;0.05. the result of RT-qPCR according to the regions of study showed that Al-shinafyah in western borders of Iraq-Saudi was the highest infection rate 35% .On the other hand ,100 human 81 nasal swabs and 19 bronchial lavage samples were collected from pilgrims and non-pilgrims. The total positive result was 5%. The pilgrims recorded the highest infection rate. The results of conventional PCR by using specific primers for detection of Nucleocapsid gene (217 bp) of MERSCoV. The results were confirmative. Three human and 11 camel positive samples were used in further sequencing and phylogenetic analysis by extraction and purification of the PCR products. Our clones sequence submitted in GenBank-NCBI for accession number.The phylogenetic tree construction and analysis results showed that most of Iraqi variants of camel and human were located in clade-B in which Saudi Arabia strains were clustered. One of our clones (MERS-Iq.2Huh) of accession number KX150500.1 was located in clade-A in the same branch of Jordanian strain while bat corona virus, SARS corona and neoromica corona virus was out group clustered in separated branch.

Epidemiology of canine distemper in Makurdi, Nigeria

Canine distemper (CD), a major disease of dogs all over the world, hitherto controlled by extensive vaccination of dogs around the world, appears to be persisting and even re-emerging in many parts of the world. It is thought that wild life reservoir hosts contribute to the emergence of CD, but in areas of the world with minimal wildlife contact with domestic dogs, it is thought that cyclical infection between clinically normal dogs and susceptible neonatal animals may be responsible for maintaining the canine distemper virus (CDV) in the canine population. We decided to examine clinically normal dogs in the Makurdi metropolis for evidence of infection with CDV, to determine if such dogs may act as sources of persistence of the CDV in the canine population. We tested 70 unvaccinated, clinically normal dogs for evidence of distemper virus using a rapid CDV antigen (Ag) chromatographic assay test kit designed for the qualitative detection of canine distemper virus antigens in urine, conjunctiva, serum or plasma. We found six (6 or 8.6%) of the 70 dogs positive for distemper antigen; three (3) of the dogs were under one (1) year of age, whereas three were 5 years or more. We conclude that the CDV is circulating among clinically normal dogs in Makurdi, and that a cyclical infection between infected but clinically normal adult dogs and puppies may be responsible for maintaining the disease in the canine population in Makurdi. Further studies are necessary to elucidate the role of vaccination and the possibilities of emergence of new antigenic strains of CDV in the epidemiology of CD in the Makurdi area. Keywords: Canine Distemper, Vaccination, Laboratory diagnosis, maintenance in population, Makurdi, Nigeria

Yellow Fever Virus: Diagnostics for a Persistent Arboviral Threat.

Yellow fever (YF) is the prototypical hemorrhagic fever and results from infection with yellow fever virus (YFV), which is endemic to regions of Africa and South America. Despite the availability of an effective vaccine, YFV continues to cause disease throughout regions where it is endemic, including intermittent large outbreaks among undervaccinated populations. A number of diagnostic methods and assays have been described for the detection of YFV infection, including viral culture, molecular testing, serology, and antigen detection. Commercial diagnostics are not widely available, and testing is generally performed at a small number of reference laboratories. The goal of this article, therefore, is to review available clinical diagnostics for YFV, which may not be familiar to many practitioners outside areas where it is endemic. Additionally, we identify gaps in our current knowledge about YF that pertain to diagnosis and describe interventions that may improve YFV detection.

A spike-modified Middle East respiratory syndrome coronavirus (MERS-CoV) infectious clone elicits mild respiratory disease in infected rhesus macaques

The recurrence of new human cases of Middle East respiratory syndrome coronavirus (MERS-CoV) underscores the need for effective therapeutic countermeasures. Nonhuman primate models are considered the gold standard for preclinical evaluation of therapeutic countermeasures. However, MERS-CoV-induced severe respiratory disease in humans is associated with high viral loads in the lower respiratory tract, which may be difficult to achieve in nonhuman primate models. Considering this limitation, we wanted to ascertain the effectiveness of using a MERS-CoV infectious clone (icMERS-0) previously shown to replicate to higher titers than the wild-type EMC 2012 strain. We observed respiratory disease resulting from exposure to the icMERS-0 strain as measured by CT in rhesus monkeys with concomitant detection of virus antigen by immunohistochemistry. Overall, respiratory disease was mild and transient, resolving by day 30 post-infection. Although pulmonary disease was mild, these results demonstrate for the first time the utility of CT imaging to measure disease elicited by a MERS-CoV infectious clone system in nonhuman primate models.

Molecular characterization of a rabies virus isolated from trade dogs in Plateau State, Nigeria

To have a better understanding of the prevalence of rabies virus (RABV) in the dog trading markets in Plateau State of Nigeria, a total of 532 brain samples collected between 2010 and 2011 were subjected to rabies virus antigen detection and molecular characterization. RABV antigen was detected in 92 out of 532 (17.3%) brain samples from two major commercial dog markets in Plateau State of Nigeria by direct fluorescence antibody assay, indicating the possibility of a high prevalence of RABV in the dog trading markets. The complete genomic sequence was obtained from one of these RABV positive samples. The overall organization of this virus (DRV-NG11) was typical of all the other wild-type RABV. Phylogenetic analysis using either the complete or partial genomic sequence of DRV-NG11 demonstrates that this isolate is most closely related to viruses previously shown to circulate in Nigeria, belonging to the Africa 2 Clade. To our knowledge, this is the first report on the molecular characterization of the complete genome of RABV from trade dogs, which provides a better understanding of the molecular epidemiology, pathogenesis and rabies control in this country.Keywords: Genome, Nigeria, Rabies virus, Street strain, Trade dogs

The structure of the C-terminal domain of the nucleoprotein from the Bundibugyo strain of the Ebola virus in complex with a pan-specific synthetic Fab.

The vast majority of platforms for the detection of viral or bacterial antigens rely on immunoassays, typically ELISA or sandwich ELISA, that are contingent on the availability of suitable monoclonal antibodies (mAbs). This is a major bottleneck, since the generation and production of mAbs is time-consuming and expensive. Synthetic antibody fragments (sFabs) generated by phage-display selection offer an alternative with many advantages over Fabs obtained from natural antibodies using hybridoma technology. Unlike mAbs, sFabs are generated using phage display, allowing selection for binding to specific strains or for pan-specificity, for identification of structural epitopes or unique protein conformations and even for complexes. Further, they can easily be produced in Escherichia coli in large quantities and engineered for purposes of detection technologies and other applications. Here, the use of phage-display selection to generate a pan-specific Fab (MJ20), based on a Herceptin Fab scaffold, with the ability to bind selectively and with high affinity to the C-terminal domains of the nucleoproteins (NPs) from all five known strains of the Ebola virus is reported. The high-resolution crystal structure of the complex of MJ20 with the antigen from the Bundibugyo strain of the Ebola virus reveals the basis for pan-specificity and illustrates how the phage-display technology can be used to manufacture suitable Fabs for use in diagnostic or therapeutic applications.

RABIES IN A SET OF EIGHT-WEEK OLD PUPPIES IN NIGERIA: THE NEED FOR REVIEW OF CURRENT DOG ANTIRABIES VACCINATION SCHEDULE.

Background:Rabies is endemic in Nigeria with clinical cases reported mainly in dogs and occasionally in livestock from all the geo-ecological zones of the country. Detection of rabies virus antigen in puppies at the age of five to ten weeks and in apparently healthy dogs shedding the virus in their saliva have been reported in some parts of Nigeria.Material and Method:This report describes a clinical rabies in a set of eight weeks old puppies confirmed by antigen detection using the direct fluorescent antibody test (DFAT), the direct rapid immunohistochemical test (DRIT), and RT-PCR.Results:it was positive for all test used including the RT-PCR which amplified at 750 bp from the gel electrophoresis.Conclusion:Occurrence of rabies in puppies of this age, within which they are acquired and owned by other unsuspecting members of the public, is of grave public health consequences. People that love puppies, especially children, who are fond of carrying and playing with them, are also faced with the risk of exposure to rabies. Consequently, review of the existing dog antirabies vaccination schedule at twelve weeks of age in Nigeria, is recommended to ensure effective immunization of this age group of dogs and for the overall safety of the vulnerable members of the public.