ABSTRACT
The removal of xenotrpic murine leukemia virus (xMuLV) by size-exclusion filter paper composed of 100% naturally derived cellulose was validated. The filter paper was produced using cellulose nanofibers derived from Cladophora sp. algae. The filter paper was characterized using atomic force microscopy, scanning electron microscopy, helium pycnometry, and model tracer (100 nm latex beads and 50 nm gold nanoparticles) retention tests. Following the filtration of xMuLV spiked solutions, LRV ≥5.25 log10 TCID50 was observed, as limited by the virus titre in the feed solution and sensitivity of the tissue infectivity test. The results of the validation study suggest that the nanocellulose filter paper is useful for removal of endogenous rodent retroviruses and retrovirus-like particles during the production of recombinant proteins.
Subject(s)
Leukemia Virus, Murine/isolation & purification , Paper , Cellulose , Drug Contamination/prevention & control , Filtration , Gold , Latex , Leukemia Virus, Murine/ultrastructure , Microscopy, Atomic Force , Microscopy, Electron, Scanning , Nanofibers , Nanospheres , Particle Size , Porosity , Solutions , Viral Load , Viral TropismABSTRACT
The pathogenesis of neurologic complications of Lassa fever is poorly understood. A Nigerian patient had fever, disorientation, seizures, and blood-brain barrier dysfunction, and Lassa virus was found in cerebrospinal fluid (CSF) but not in serum. The concentration of Lassa virus RNA in CSF corresponded to 1 x 10(3) pfu/mL, as determined by a quantitative real-time polymerase chain reaction assay. To characterize the Lassa virus in CSF, the 3.5-kb S RNA was sequenced. In the S RNA coding sequences, the CSF strain differed between 20% and 24.6% from all known prototype strains. These data suggest that Lassa virus or specific Lassa virus strains can persist in the central nervous system and thus contribute to neuropathogenesis. Lassa virus infection should be considered in West African patients or in travelers returning from this area who present only with fever and neurologic signs.
Subject(s)
Confusion/etiology , Lassa Fever/cerebrospinal fluid , Lassa Fever/complications , Lassa virus/genetics , Lassa virus/isolation & purification , Seizures/etiology , Amino Acid Sequence , Confusion/cerebrospinal fluid , Confusion/virology , Fever/etiology , Humans , Lassa Fever/blood , Lassa virus/classification , Male , Middle Aged , Molecular Sequence Data , Nigeria , Phylogeny , Polymerase Chain Reaction/methods , RNA, Viral/chemistry , RNA, Viral/genetics , Reverse Transcriptase Polymerase Chain Reaction , Seizures/cerebrospinal fluid , Seizures/virology , Sequence Alignment , Sequence Homology, Amino AcidABSTRACT
Callitrichid hepatitis (CH) is a highly fatal, rodent-borne zoonosis of New World primates (family Callitrichidae) caused by lymphocytic choriomeningitis virus (LCMV). It is unclear whether virulence in Callitrichidae is associated with specific genetic or phylogenetic markers of the virus as only a partial S RNA sequence of a single CH-associated isolate is known. In a period of 10 months, three pygmy marmosets (Cebuella pygmaea) and one Goeldi's monkey (Callimico goeldii) died from CH in a German zoo. LCMV was most likely transmitted by wild mice. Infection was associated with characteristic histopathological lesions in liver, brain, and lymphoid tissue. Virus sequences from all callitrichids and a captured mouse were > or =99.2% identical. LCMV strains from a pygmy marmoset and the Goeldi's monkey were isolated in cell culture and the 3.4-kb S RNA was completely sequenced. Both strains differed considerably in their genetic and phylogenetic characteristics from known LCMV strains, including the previously described CH-associated strain. These data show that CH is widespread and can be caused by distantly related LCMV strains.
Subject(s)
Animals, Zoo/virology , Callimico/virology , Callitrichinae/virology , Disease Outbreaks/veterinary , Genome, Viral , Hepatitis, Viral, Animal/virology , Lymphocytic choriomeningitis virus/isolation & purification , Monkey Diseases/virology , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Fluorescent Antibody Technique , Germany , Hepatitis, Viral, Animal/epidemiology , Humans , L Cells , Liver/pathology , Liver/virology , Lymphocytic choriomeningitis virus/genetics , Male , Mice , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , RNA, Viral/analysis , Sequence AlignmentABSTRACT
We describe the isolation and characterization of a new Lassa virus strain imported into Germany by a traveler who had visited Ghana, Côte D'Ivoire, and Burkina Faso. This strain, designated "AV," originated from a region in West Africa where Lassa fever has not been reported. Viral S RNA isolated from the patient's serum was amplified and sequenced. A long-range reverse transcription polymerase chain reaction allowed amplification of the full-length (3.4 kb) S RNA. The coding sequences of strain AV differed from those of all known Lassa prototype strains (Josiah, Nigeria, and LP) by approximately 20%, mainly at third codon positions. Phylogenetically, strain AV appears to be most closely related to strain Josiah from Sierra Leone. Lassa viruses comprise a group of genetically highly diverse strains, which has implications for vaccine development. The new method for full-length S RNA amplification may facilitate identification and molecular analysis of new arenaviruses or arenavirus strains.
