These results confirmed that this Cambodian isolates are closely related to the other NiV isolates. Phylogenetic analysis using parsimony method (subfamily confirmed that this Cambodian isolates and the Malaysian NiV isolates were significantly comparable and that the Cambodian strains probably belong to the NiV species (Figure 2). from June 2003 to August 2004 on plastic linens, following a published procedure (or specific primers for the nucleoprotein (N) gene of NiV (were positive (4 with a titer = 1:10 and 2 with a titer = 1:20). The 108 ELISA-negative serum samples from other captured species, (n = 15), (n = 15), (n = 15), (n = 27), (n = 12), and (n = 24), were negative. None of the human sera were NiV seropositive with the seroneutralization test. Computer virus Isolation and Molecular Characterization Attempts to isolate viruses producing a CPE were positive for 2 samples among the 769 urine samples. These 2 samples, CSUR381 and CSUR382, were collected consecutively on the same morning (April 23, 2004) at the same roost of specimens, located in a pagoda in the village of Bay Damran in Battambang Province. CPE showing syncytia was detected 6 Vorinostat (SAHA) days after injection into Vero E6 cells. RT-PCR performed on RNA extracted from infected cell Mouse monoclonal to CD86.CD86 also known as B7-2,is a type I transmembrane glycoprotein and a member of the immunoglobulin superfamily of cell surface receptors.It is expressed at high levels on resting peripheral monocytes and dendritic cells and at very low density on resting B and T lymphocytes. CD86 expression is rapidly upregulated by B cell specific stimuli with peak expression at 18 to 42 hours after stimulation. CD86,along with CD80/B7-1.is an important accessory molecule in T cell costimulation via it’s interaciton with CD28 and CD152/CTLA4.Since CD86 has rapid kinetics of induction.it is believed to be the major CD28 ligand expressed early in the immune response.it is also found on malignant Hodgkin and Reed Sternberg(HRS) cells in Hodgkin’s disease supernatants using specific primers for the P gene of isolate). We observed that this Cambodian N nucleotide sequence shared 98% identity with the Malaysian AF21232 N sequence (32 nucleotides among 1,599 were divergent). The identity was 98.7% (525/532) for the N amino acid sequences (Figure A1). All the changes of the 532 amino acid (aa) N protein occurred at the carboxyl terminus, with the following mutations: I429V, G432E, N457D, I502T, E511G, L518P, and A521T. The G sequence of the Cambodian CSUR381 strain was also compared to the available G sequences of the Malaysian NiV isolates. The percentage of nucleotide homology was 98.2% (32 nucleotide changes among 1,809), and the percentage of amino acid homology was 98.5% (593/602). Amino acid changes were N5S, V24I, R248K, G327D, I408V, V426I, L470Q, N478S, and N481D (Physique A2). These results confirmed that this Cambodian isolates are closely related to the other NiV isolates. Phylogenetic analysis using parsimony method (subfamily confirmed that this Cambodian isolates and the Malaysian NiV isolates were significantly comparable and that the Cambodian strains probably belong to the NiV species (Physique 2). Analyses performed by using other methods (neighbor joining and maximum likelihood) or using the G sequences reached the same conclusion (data not shown). The N and G nucleotide sequences of the isolate CSUR381 were deposited in GenBank (accession nos. “type”:”entrez-nucleotide”,”attrs”:”text”:”AY858110″,”term_id”:”61676705″,”term_text”:”AY858110″AY858110 and “type”:”entrez-nucleotide”,”attrs”:”text”:”AY858111″,”term_id”:”61676707″,”term_text”:”AY858111″AY858111). Open in a separate window Physique 2 Phylogenetic analysis of the 1,599 nucleotides of the N gene coding domain name sequence from the Nipah computer virus Cambodian isolate, members of the subfamily used as outgroups. GenBank accessions numbers used are as follows: APMV-6: Avian paramyxovirus 6, “type”:”entrez-nucleotide”,”attrs”:”text”:”AY029299″,”term_id”:”15072442″,”term_text”:”AY029299″AY029299; CDV: Canine distemper virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF014953″,”term_id”:”3335048″,”term_text”:”AF014953″AF014953; DMV, Dolphin distemper computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X75961″,”term_id”:”557234″,”term_text”:”X75961″X75961; HeV: Hendra computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF017149″,”term_id”:”529283688″,”term_text”:”AF017149″AF017149; HPIV-1: Human parainfluenza computer virus 1, D011070; HPIV-2: Human parainfluenza computer virus 2, “type”:”entrez-nucleotide”,”attrs”:”text”:”M55320″,”term_id”:”332736″,”term_text”:”M55320″M55320; HPIV-3: Human parainfluenza computer virus 3, “type”:”entrez-nucleotide”,”attrs”:”text”:”D10025″,”term_id”:”3327816″,”term_text”:”D10025″D10025; HPIV4a: Parainfluenza computer virus type 4A, “type”:”entrez-nucleotide”,”attrs”:”text”:”M32982″,”term_id”:”332581″,”term_text”:”M32982″M32982; HPIV4b: Parainfluenza computer virus type 4B, “type”:”entrez-nucleotide”,”attrs”:”text”:”M32983″,”term_id”:”332583″,”term_text”:”M32983″M32983; HRSV: Human respiratory syncytial computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X00001″,”term_id”:”61215″,”term_text”:”X00001″X00001; MeV: Vorinostat (SAHA) Measles computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”K01711″,”term_id”:”331784″,”term_text”:”K01711″K01711; MPRV: Mapuera computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X85128″,”term_id”:”1019793″,”term_text”:”X85128″X85128; Menangle: Menangle computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF326114″,”term_id”:”82393544″,”term_text”:”AF326114″AF326114; Mossman: Mossman computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AY286409″,”term_id”:”33637703″,”term_text”:”AY286409″AY286409; MuV: Mumps computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”D86172″,”term_id”:”1468930″,”term_text”:”D86172″D86172; Vorinostat (SAHA) NDV: Newcastle disease computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF064091″,”term_id”:”3139149″,”term_text”:”AF064091″AF064091; NiV: Nipah computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF212302″,”term_id”:”13518006″,”term_text”:”AF212302″AF212302; PDV: Phocid distemper computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X75717″,”term_id”:”440568″,”term_text”:”X75717″X75717; PPRV, Peste des Petits ruminants computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X74443″,”term_id”:”67906091″,”term_text”:”X74443″X74443; RPV: Rinder pest computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X68311″,”term_id”:”61933″,”term_text”:”X68311″X68311; Salem: Salem computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF237881″,”term_id”:”7546986″,”term_text”:”AF237881″AF237881; SeV: Sendai computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”X00087″,”term_id”:”60928″,”term_text”:”X00087″X00087; SV5: Simian computer virus 5, “type”:”entrez-nucleotide”,”attrs”:”text”:”M81442″,”term_id”:”335114″,”term_text”:”M81442″M81442; TiV: Tioman computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF298895″,”term_id”:”21930305″,”term_text”:”AF298895″AF298895; TPMV: Tupaia paramyxovirus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AF079780″,”term_id”:”7536530″,”term_text”:”AF079780″AF079780; TRTV: Turkey Rhinotracheitis computer virus, “type”:”entrez-nucleotide”,”attrs”:”text”:”AY640317″,”term_id”:”49823134″,”term_text”:”AY640317″AY640317. Significant bootstrap values (70%) are indicated. The phylogram was generated by parsimony method and analyzing 100 bootstrap replicates. Discussion Our serologic study confirms the presence in Cambodia of antibodies to a NiV-like computer virus among Lyle’s Flying Foxes (and have been found carrying neutralizing antibodies to NiV in Malaysia. These specimens were collected soon after an outbreak, when the computer virus was expected to circulate with high prevalence ((is the first isolate obtained from a species different from (family is the most conserved region, which seems to be involved in its interactions with other functional proteins, such as the phosphoprotein (P) and the polymerase (L). Moreover the N proteins of paramyxoviruses possess 3 highly conserved regions (Reynes JM, Counor D, Ong S, Faure C, Seng V, Molia.