A VLP-based vaccine provides complete protection against Nipah virus challenge following multiple-dose or single-dose vaccination schedules in a hamster model.

Nipah virus is a highly lethal zoonotic paramyxovirus that was first recognized in Malaysia during an outbreak in 1998. During this outbreak, Nipah virus infection caused a severe febrile neurological disease in humans who worked in close contact with infected pigs. The case fatality rate in humans was approximately 40%.

Mammalian Cell-Derived Respiratory Syncytial Virus-Like Particles Protect the Lower as well as the Upper Respiratory Tract.

Globally, Respiratory Syncytial Virus (RSV) is a leading cause of bronchiolitis and pneumonia in children less than one year of age and in USA alone, between 85,000 and 144,000 infants are hospitalized every year. To date, there is no licensed vaccine. We have evaluated vaccine potential of mammalian cell-derived native RSV virus-like particles (RSV VLPs) composed of the two surface glycoproteins G and F, and the matrix protein M.

Vaccine potential of Nipah virus-like particles.

Nipah virus (NiV) was first recognized in 1998 in a zoonotic disease outbreak associated with highly lethal febrile encephalitis in humans and a predominantly respiratory disease in pigs. Periodic deadly outbreaks, documentation of person-to-person transmission, and the potential of this virus as an agent of agroterror reinforce the need for effective means of therapy and prevention. In this report, we describe the vaccine potential of NiV virus-like particles (NiV VLPs) composed of three NiV proteins G, F and M.

Cis-acting elements in the antigenomic promoter of Nipah virus.

Genome synthesis in paramyxoviruses, including Nipah virus (NiV), is controlled by sequence elements that reside in the non-coding nucleotides at the 5'-trailer (3'-antigenomic) end that make up the antigenomic promoter (AGP). Using a chloramphenicol acetyl transferase-based plasmid-driven minigenome system, the terminal 96 nt of NiV AGP were first mutagenized in blocks of three hexamers to enable broad mapping of the minigenome functional regions. This was followed by further dissection of these functional regions to define the cis-acting elements contained therein.

Reverse genetics of negative-stranded RNA viruses: a global perspective.

The advent of reverse genetics technology has revolutionized the field of RNA viruses. It is now possible to manipulate even negative-stranded RNA viruses at will, and evaluate the effects of these changes on the biology and pathogenesis of these viruses. The fundamental insights gleaned from the reverse genetics-based studies over the last several years have provided a new momentum for the development of designed therapies for the control and prevention of these viral pathogens.

An internal element of the measles virus antigenome promoter modulates replication efficiency.

The cis-acting sequence elements that direct measles virus (MV) genome synthesis reside in the 109 base non-coding region at the 5' trailer (3' antigenomic) end of MV genome that makes up the antigenomic promoter (AGP). The MV-AGP nucleotides 79-96, corresponding to nucleotide hexamers 14, 15 and 16 (the C' element), show sequence similarity with the equivalent region of many paramyxoviruses and are analogous to the three nucleotide hexamers that form the second replication control element in the Sendai virus AGP. In this study, results of two independent procedures demonstrate that the MV C' element also is a replication control sequence.

Expression of a foreign gene by recombinant canine distemper virus recovered from cloned DNAs.

A canine distemper virus (CDV) genomic cDNA clone and expression plasmids required to establish a CDV rescue system were generated from a laboratory-adapted strain of the Onderstepoort vaccine virus. In addition, a CDV minireplicon was prepared and used in transient expression studies performed to identify optimal virus rescue conditions. Results from the transient expression experiments indicated that minireplicon-encoded reporter gene activity was increased when transfected cell cultures were maintained at 32 rather than 37 degrees C, and when the cellular stress response was induced by heat shock.

Analysis of the noncoding regions of measles virus strains in the Edmonston vaccine lineage.

The noncoding sequence of five Edmonston vaccine viruses (AIK-C, Moraten, Rubeovax, Schwarz, and Zagreb) and those of a low-passage Edmonston wild-type (wt) measles virus have been determined and compared. Twenty-one nucleotide positions were identified at which Edmonston wt and one or more vaccine strains differed. The location of some of these nucleotide substitutions suggests that they may influence the efficiency of mRNA synthesis, processing, and translation, as well as genome replication and encapsidation.

Comparison of predicted amino acid sequences of measles virus strains in the Edmonston vaccine lineage.

Protein-encoding nucleotide sequences of the N, P, M, F, H, and L genes were determined for a low-passage isolate of the Edmonston wild-type (wt) measles virus and five Edmonston-derived vaccine virus strains, including AIK-C, Moraten, Schwarz, Rubeovax, and Zagreb. Comparative analysis demonstrated a high degree of nucleotide sequence homology; vaccine viruses differed at most by 0. 3% from the Edmonston wt strain.

Enhanced measles virus cDNA rescue and gene expression after heat shock.

Rescue of negative-stranded RNA viruses from full-length genomic cDNA clones is an essential technology for genetic analysis of this class of viruses. Using this technology in our studies of measles virus (MV), we found that the efficiency of the measles virus rescue procedure (F. Radecke et al.

Detection of Bordetella pertussis and respiratory synctial virus in air samples from hospital rooms.

Objective: To evaluate the distribution of Bordetella pertussis and respiratory syncytial virus (RSV) in the hospital setting.

Design: Air samples were collected using filters in the hospital rooms of 12 children with pertussis and 27 children with RSV infection. Material eluted from these filters was subjected to RSV- and B pertussis-specific polymerase chain reaction (PCR) amplification.

Mammalian epithelial cell line kit for detection of Clostridium difficile toxin.

The performance characteristics of a mammalian epithelial (MEP) cell line kit (Cytotoxi Test; Advanced Clinical Diagnostics, Toledo, Ohio) for the detection of Clostridium difficile toxin was compared with that of conventional tissue culture assays with human embryonic lung (HEL) cells in shell vials and human foreskin fibroblasts (HFFs) in test tubes. One hundred forty-nine stool samples were tested. The MEP cells were at least as sensitive as the HEL cells for use in C.

A review of sudden infant death syndrome.

This chapter defines sudden infant death syndrome (SIDS) and provides a brief review of the history, current knowledge, pathology, and possible mechanisms of death. The authors conclude with considerations of other selected causes of sudden, unexpected infant death.

Distinguishing between respiratory syncytial virus subgroups by protein profile analysis.

We subgrouped 75 strains of respiratory syncytial virus by a protein profile method (PPM) which relies on different mobilities of the phosphoprotein in one-dimensional polyacrylamide gel electrophoresis and does not require monoclonal antibodies. When compared with enzyme immunoassay, PPM correctly subgrouped 54 of 56 subgroup A and all 19 subgroup B strains.

Rapid detection of respiratory syncytial virus in nasopharyngeal specimens obtained with the rhinoprobe scraper.

We compared the Rhinoprobe scraping technique for collection of superficial nasal mucosa epithelial cells and rapid detection of respiratory syncytial virus by immunofluorescence with paired, swab-collected specimens for virus culture from 1,257 infants and children with acute respiratory infections. Compared with viral culture as the reference test, the sensitivity, specificity, and accuracy of the immunofluorescence test were 83.6, 93.

Protein fingerprinting: a novel virus identification system.

Viral proteins separated by one-dimensional SDS-PAGE produce protein binding patterns (fingerprints) which are unique for different viruses. We have applied this concept successfully for the development of a practical and objective virus identification system which is applicable to most viruses. The method is simple, specific, and, unlike the currently available methods, free from all virus-specific reagents.

Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies.

Identification of herpes simplex viruses by automated profile analysis of viral proteins.

In this report we describe a computerized virus identification system based upon the fact that viral proteins, separated by one-dimensional (ID) SDS-PAGE have unique banding patterns (protein profiles). Fifty-eight clinical strains of herpes simplex virus (HSV) were correctly identified by comparing their protein profiles with reference profiles of HSV, other viruses and uninfected cell cultures stored in a computer data base.

Dual infection of the conjunctiva with herpes simplex virus and Chlamydia trachomatis.

We describe a homosexual man with simultaneous infection of the conjunctiva by Herpes simplex virus and Chlamydia trachomatis. This dual infection was associated with a genital and disseminated Herpes simplex virus infection as well as asymptomatic chlamydial infection of the rectum and "nonspecific" urethritis. The findings in this case show the importance of laboratory investigation in cases of conjunctivitis associated with genital infection.

Value and cost effectiveness of double culture tests for diagnosis of ocular viral and chlamydial infections.

Swabbings from the eyes of 4132 patients attending ophthalmic casualty and outpatients clinics were tested for chlamydiae, adenovirus, and herpes simplex virus. Laboratory isolation tests gave positive results for one of these three agents in 696 (16.8%) cases.

Development of an immunofluorescence test for the serodiagnosis of herpes zoster ophthalmicus.

An indirect immunofluorescence test has been developed and evaluated for the serodiagnosis of herpes zoster ophthalmicus (HZO) by the detection of antivaricella zoster virus (VZV) antibody. The results show that, in patients with HZO, anti-VZV IgG antibody titre usually rises rapidly after onset. One hundred and seven of the 134 sera (80%) from patients with a clinical diagnosis of HZO had an anti-VZV IgG titre of greater than or equal to 256, and IgM antibody at a level of 1 in 8 was present in six of them.

A rapid and sensitive culture test for the laboratory diagnosis of genital herpes in women.

A rapid and sensitive cell culture test has been developed to detect herpes simplex virus (HSV) in women with genital herpes. The virus is cultured by inoculation and centrifugation of cell monolayers, and the virus inclusions are detected using an indirect immunofluorescence test. The test takes only 48 hours to complete compared with the conventional cell culture test, which may take up to eight days.

A rapid and sensitive culture test for detecting herpes simplex virus from the eye.

A rapid and sensitive culture test has been developed for detecting herpes simplex virus (HSV) in ocular infections. The virus is cultured by inoculation and centrifugation of cell monolayers grown on coverslips and the inclusions detected by an indirect immunofluorescence technique. This rapid test takes only two days to complete.

Rapid culture test for adenovirus isolation.

A rapid culture test has been developed and evaluated for the detection of adenovirus in ocular infections. The test requires only two days' incubation of the inoculated cell monolayers on coverslips in flat bottomed tubes followed by detection of adenovirus inclusions using immunofluorescence staining method. The sensitivity of the rapid test is found to be comparable with that of the conventional tissue culture test which depends on the development of a cytopathic effect requiring 2-21 days' incubation (mean 9 X 4, SD 4 X 9).