title: RT-PCR assay for detection of Lassa virus and related Old World arenaviruses targeting the L gene.
abstract: This study describes an RT-PCR assay targeting the L RNA segment of arenaviruses. Conserved regions were identified in the polymerase domain of the L gene on the basis of published sequences for Lassa virus, lymphocytic choriomeningitis virus (LCMV), Pichinde virus and Tacaribe virus, as well as 15 novel sequences for Lassa virus, LCMV, Ippy virus, Mobala virus and Mopeia virus determined in this study. Using these regions as target sites, a PCR assay for detection of all known Old World arenaviruses was developed and optimized. The concentration that yields 95% positive results in a set of replicate tests (95% detection limit) was determined to be 4290 copies of Lassa virus L RNA per ml of serum, corresponding to 30 copies per reaction. The ability of the assay to detect various Old World arenaviruses was demonstrated with in vitro transcribed RNA, material from infected cell cultures and samples from patients with Lassa fever and monkeys with LCMV-associated callitrichid hepatitis. The L gene PCR assay may be applicable: (i) as a complementary diagnostic test for Lassa virus and LCMV; (ii) to identify unknown Old World arenaviruses suspected as aetiological agents of disease; and (iii) for screening of potential reservoir hosts for unknown Old World arenaviruses.
authors: Department of Virology, Bernhard-Nocht-Institute for Tropical Medicine, Bernhard-Nocht-Strasse 74, D-20359 Hamburg, Germany.
authors: Simon
authors: S
authors: Vieth
authors: Christian
authors: C
authors: Drosten
authors: Oliver
authors: O
authors: Lenz
authors: Martin
authors: M
authors: Vincent
authors: Sunday
authors: S
authors: Omilabu
authors: Meike
authors: M
authors: Hass
authors: Beate
authors: B
authors: Becker-Ziaja
authors: Jan
authors: J
authors: ter Meulen
authors: Stuart T
authors: ST
authors: Nichol
authors: Herbert
authors: H
authors: Schmitz
authors: Stephan
authors: S
authors: Günther
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001117:Arenaviridae Infections / Q000175:diagnosis* / Q000235:genetics; D018049:Arenaviruses, Old World / Q000145:classification / Q000235:genetics; D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis; D000882:Haplorhini; D006801:Humans; D007835:Lassa Fever / Q000175:diagnosis* / Q000235:genetics; D007836:Lassa virus / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral