The complete nucleotide sequence of isolate BYMV-GDD of <Emphasis Type="Italic">Bean yellow mosaic virus</Emphasis>, and comparison to other potyviruses

Summary. The complete nucleotide sequence of Bean yellow mosaic virus (BYMV) gladiolus isolate GDD was determined and compared to broad bean isolates BYMV-MB4 and BYMV-S. The BYMV-GDD genome (9528nt) was more similar to BYMV-MB4 (9532nt) than to BYMV-S (9547nt), which has atypical symptom expression and host range. The greatest variability occurred in the 5 untranslated region, P1 protein, and NIa-VPg protein, the N-terminal two thirds of HC-Pro, and the C-terminal one third of P3. Each of these regions has been correlated with symptom or host differences between isolates of other potyviruses, and may contribute to the atypical nature of BYMV-S.     

Mother-Baby psychiatric units (MBUs): National data collection in France and in Belgium (1999–2000)

Summary The French version of the Marcé checklist was used to collect data for 176 joint admissions to 11 psychiatric mother-baby units in 1999 and 2000. Mean age of the babies at admission ranged from 4 to 16 weeks. Two units also admitted older children. Mothers admitted were diagnosed with schizophrenia or chronic delusional disorders (n=44), acute transitory psychosis Bouffée délirante (n=20), bipolar disorders (n=20), depressive illness (n=38), personality disorders or intellectual disability (n=39), and other disorders (n=15). The mean duration of hospitalisation was 11 weeks. Units that also offered day-care admission in the same or a near-by unit had shorter mean admissions. More than half the womens partners (or babies fathers) had mental health problems. Women with schizophrenia or chronic delusional disorders and personality disorders or intellectual disability remained hospitalised longer, improved less, and were more often separated from their babies, or discharged with supervision, than women admitted with other diagnoses.     

The complete nucleotide sequence of the genome RNA of Lily symptomless virus and its comparison with that of other carlaviruses

Summary. The complete genomic nucleotide sequence and genome structure of Lily symptomless virus (LSV), a lily-infecting carlavirus, have been obtained. The genome of the Korean strain of LSV, LSV-Kr, was 8,394 nucleotides long and contained six open reading frames (ORFs) coding for proteins of Mr 220kDa (1,948aa), 25kDa (228aa), 12kDa (106aa), 7kDa (64aa), 32kDa (291aa) and 16kDa (140aa) from the 5 to 3 end, respectively, which is typical of carlaviruses. Genetic heterogeneity was observed in the ORF1 gene. A total of 221 of 5,847 nucleotides (nt) were heterologous in the ORF1 of replicase; 162nt portions were silent and 59nt resulted in amino acid changes. This heterogeneity indicates that the LSV-infecting lily plants contained a genetically heterogeneous population of LSV (quasispecies). Overall similarities to those of other carlaviruses for the six ORFs of LSV were from 76.1% to 31.6% and from 87.3% to 13.7%, at nucleotide and amino acid levels, respectively. The ORF1 replicase gene of LSV shares 40.9% to 56.8% and 48.9% and 58.6% identities with that of 5 other carlaviruses at the amino acid and nucleotide levels, respectively. LSV was closest to Blueberry scorch virus (BlScV) in this ORF, among the carlaviruses for which sequence information is available. The three triple gene blocks (ORF2-4), ORF5 (coat protein) and 3-proximal 16kDa ORF6 genes were further analyzed, and phylogenetic trees for the coding regions indicate that the LSV was the most closely related to Kalanchoe latent virus and BlScV. This is the first report of the complete nucleotide sequence and genome structure of LSV.Received December 13, 2002; accepted May 14, 2003 Published online July 17, 2003     

Changes in level of virus accumulation and incidence of infection are critical in the characterization of <Emphasis Type="Italic">Rice tungro bacilliform virus</Emphasis> (RTBV) resistance in rice

Summary. Analysis by enzyme-linked immunosorbent assay showed that Rice tungro bacilliform virus (RTBV) accumulated in a cyclic pattern from early to late stages of infection in tungro-susceptible variety, Taichung Native 1 (TN1), and resistant variety, Balimau Putih, singly infected with RTBV or co-infected with RTBV+Rice tungro spherical virus (RTSV). These changes in virus accumulation resulted in differences in RTBV levels and incidence of infection. The virus levels were expressed relative to those of the susceptible variety and the incidence of infection was assessed at different weeks after inoculation. At a particular time point, RTBV levels in TN1 or Balimau Putih singly infected with RTBV were not significantly different from the virus level in plants co-infected with RTBV+RTSV. The relative RTBV levels in Balimau Putih either singly infected with RTBV or co-infected with RTBV+RTSV were significantly lower than those in TN1. The incidence of RTBV infection varied at different times in Balimau Putih but not in TN1, and to determine the actual infection, the number of plants that became infected at least once anytime during the 4wk observation period was considered. Considering the changes in RTBV accumulation, new parameters for analyzing RTBV resistance were established. Based on these parameters, Balimau Putih was characterized having resistance to virus accumulation although the actual incidence of infection was >75%.Received December 18, 2002; accepted March 19, 2003 Published online June 5, 2003     

Actin filaments around endothelial fenestrae in rat hepatic sinusoidal endothelial cells

The presence of microfilaments in the vicinity of sinusoidal endothelial fenestrae (SEF) suggests that the cytoskeleton of liver sinusoidal endothelial cells (LSEC) plays an important role in the modulation of SEF. In this study, we investigated actin filaments around SEF in LSECs. Monolayers of LSEC culture were established by infusing a rat liver with collagenase for 30min and then culturing in RMPI medium for 24h. Cells were reacted with 0.1% Triton X for 5s and 15% glycerinated PHEM buffer (60mM PIPES, 25mM HEPES, 10mM EGTA, 2mM MgCl, pH 6.9) containing heavy meromyosin for 10min and observed under a transmission electron microscope. By electron microscopy with the modified heavy meromyosin decorated reaction, actin filaments were clearly demonstrated around SEF in LSEC.     

Synthesis of Seoul virus RNA and structural proteins in cultured cells

Summary. Seoul virus is a hantavirus that causes hemorrhagic fever with renal syndrome (HFRS). The virion has a tripartite (S, M, and L) negative-stranded RNA genome, which is characteristic of the family Bunyaviridae. However, the molecular basis of virus replication is not well known. We established a Northern blot hybridization (NB) procedure using digoxygenin-labeled RNA probes, to quantitate the hantaviral plus- and minus-strand RNAs separately. Virus RNA replication was analyzed in infected Vero E6 cells. When the Vero E6 cells were infected with Seoul virus strain KI-83-262 (KI) at m.o.i.=0.25, the plus-strand RNA was detected within 1h post-infection (hpi), and the minus-strand RNA was detected subsequently. Using laser confocal microscopy, the nucleocapsid protein (NP) was detected within 2hpi, and accumulated as scattered granules in the cytoplasm until 24hpi. In contrast, the G2 protein first appeared at 8hpi, was immediately transported to the Golgi, and accumulated in the Golgi until 24hpi. Infectious virus particles were released into the medium at 24hhpi. These findings indicate that hantavirus RNA replication starts with the appearance of NP at 2hpi, glycoproteins then accumulate gradually in the Golgi, and virion formation is initiated once the viral RNAs and proteins have accumulated.Received October 10, 2002; accepted April 25, 2003 Published online July 17, 2003     

Molecular analysis of <Emphasis Type="Italic">Fiji disease virus</Emphasis> genome segments 5, 6, 8 and 10

Summary. The complete sequences of Fiji disease virus (FDV) genome segments 5 (S5), S6, S8 and S10 were obtained and comprised 3150nt, 2831nt, 1959nt and 1819nt, respectively. Each segment contained a single ORF which encoded putative proteins of 115kDa, 97kDa, 69kDa and 63.0kDa, respectively. The putative amino acid sequences encoded by S5 and S6 contained putative leucine zipper motifs while FDV S5 and S8 each contained an ATP-GTP-binding motif. At the amino acid level, FDV S5, S6, S8 and S10 showed most similarity to the corresponding segments of Rice black-streaked dwarf virus. Based on sequence similarities, it is predicted that FDV S8 encodes a minor core protein, while FDV S10 encodes an outer capsid protein. The evolutionary relationships of FDV to other reoviruses are discussed.The nucleotide sequence data for FDV S5, S6, S8 and S10 are available in the DDBJ/EMBL/GenBank databases under the accession numbers AY029521, AF356083, AY297693 and AY297694, respectively.     

A temperature-dependent inhibitory activity of serum on the capacity of Saccharomyces cerevisiae-derived hepatitis B surface antigen to bind to monocytes

Summary. Hepatitis B surface antigen, when produced in yeast (rHBsAg), is capable of binding to cells that express the lipopolysaccharide coreceptor CD14. This interaction is enhanced by a serum protein, the lipopolysaccharide binding protein (LBP). Here we report that most of the rHBsAg particles that attached to monocytes at 0°C, were not endocytosed but were released back into the serum-containing binding buffer at 37°C. Additionally, serum-dependent binding at 37°C was weak when compared to the serum-dependent attachment at 0°C. Pre-incubation at 37°C of cells together with serum did not abolish binding of freshly added rHBsAg at 0°C. However, pre-incubation of rHBsAg with serum at 37°C reduced attachment to cells following incubation at 0°C. Soluble CD14 and LBP, two serum proteins which can act as phospholipid transfer molecules, were shown not to be responsible for the inhibitory effect. Pre-incubation at 37°C of rHBsAg in serum-free hepatoma cell line-conditioned media resulted in a pronounced reduction in subsequent binding to cells at 0°C. These observations suggest that the temperature-dependent inhibitory effect is caused by serum factors that are probably secreted by hepatocytes.     

Sequence analysis of the fragment of the phosphoprotein gene of Polish distemper virus isolates

Summary. The nucleotide sequence analysis of the 429bp fragment of the P gene of 11 Polish field isolates of Canine distemper virus (CDV), reference strains and other virus isolates available in the GenBank was the aim of the studies. High homology between all dog strains from east-southern region of Poland and reference strains of CDV was demonstrated. It was estimated as 97–100% for CDV-OND; 96.7–99.8% for CDV-Rock; 96.7–99.8% for CDV-LED and 96.3–97.9% for A75-17. The 100% homology of the nucleotide sequence was observed between CDV Pulawy 92, CDV Pulawy 97 and the reference CDV-OND. The homology between CDV-OND and viruses isolated from the mink and ferret was estimated as 97.7% and 98.4%, respectively. Virus strains isolated from blue foxes demonstrated the highest homology to CDV-OND – equal to 97.7% for DV 79 and 99.5% for DV 92. The fox isolate from 1992 had higher level of homology to dog isolates (96.5–99.5%) than the strain isolated from the fox in 1979 (97.2–98.8%). The phylogenetic tree has two main lineages representing two separated genetic groups: I containing PDV and II containing all distemper virus strains isolated from terrestrial carnivores. CDV strains isolated from dogs from Pulawy region between 1992–1998 and from the fox (DV 92) formed the separate lineage containing also reference strains. They differed from the native isolates from the mink and ferret as well as from Japanese strains of CDV.Received October 29, 2002; accepted April 2, 2003 Published online June 11, 2003     

Analysis of cytosolic and lysosomal pH in apoptotic cells by flow cytometry

Several reports indicate that the cytosol is acidified during apoptosis although the mechanism is not yet fully elucidated. The most acidic organelle found in the cell is the lysosome, raising the possibility that lysosomal proton release may contribute to the cytosolic acidification. We here describe methods for measurement of the cytosolic and lysosomal pH in U937 cells by a dual-emission ratiometric technique suitable for flow cytometry. Cytosolic pH was analysed in cells loaded with the fluorescent probe BCECF, while lysosomal pH was determined after endocytosis of FITC-dextran. Standard curves were obtained by incubating cells in buffers with different pH in the presence of the proton ionophore nigericin. Apoptosis was induced by exposure of cells to 10ng/ml TNF- for 4h, and apoptotic cells were identified using a fluorescent marker for active caspases. By gating of control and apoptotic cells, the cytosolic and lysosomal pH were calculated in each population. The cytosolic pH was found to decrease from 7.2 ± 0.1 to 5.8s±0.1 and the lysosomal increased from 4.3±0.4 to 5.2±0.3. These methods will be useful in future attempts to evaluate the involvement of lysosomes in the acidification of the cytosol during apoptosis.     

Detection of enteric viruses and bacterial indicators in German environmental waters

Summary. A German mining lake and the supplying surface waters, which are located downstream of a sewage plant, were examined regarding their microbiological and virological quality. Between October 2002 and September 2003, specific PCR methods were used to determine the occurrence of enteric viruses in 123 water specimens drawn at different sites downstream of the waste water treatment plant and in 9 samples from the sewage plant influent. Detection rates in sewage plant effluents and surface water samples depended on sampling sites and were: 29–76% for enterovirus (EntV), 24–42% (astrovirus, AstV), 15–53% (norovirus, NV), 3–24% (rotavirus, RoV), 5–20% (hepatitis A virus, HAV) and 20% (adenovirus, AdV). AstV genome load of selected samples was between 3.7×10³ to 1.2×10⁸ genome equivalents per liter (gen.equ./l), depending on sampling location; NV average genome load ranged from 1.8×10⁴ to 9.7×10⁵gen.equ./l. Cell culture methods showed that three out of 18 PCR positive samples contained infectious EntV. Even though microbiolical parameters such as Escherichia coli, enterococci and coliphages indicated acceptable microbiological water quality, the virological data of this study suggest the possibility that surface waters may be a source for enteric viral infections.     

Development of a dialyzer with enhanced internal filtration to increase the clearance of low molecular weight proteins

Accumulated low molecular weight proteins in hemodialysis patients require a high-flux dialyzer. There have been several methods proposed for enhancing internal filtration, including narrowing the inside diameter of the hollow fibers, lengthening the fibers, and increasing the fiber density ratio. We tried to enhance the internal filtration by increasing the pressure drop in the dialysate compartment through increasing the fiber density ratio. If the fiber density ratio is too high, however, an irregular dialysate path may result, thus decreasing dialysis performance. Therefore, we took note of the shape of the inner housing and added a short taper structure, which improved the dialysate path dramatically. Consequently, we developed an internal filtration-enhanced dialyzer (APS-Prototype) to improve dialysis performance. The internal filtration rate in water (measured by Doppler ultrasound) was 13.2l/session for the APS-Prototype and 5.3l/session for the APS-15E. The amount of 1-microglobulin (1-MG) in bovine plasma was 0.34g for the APS-Prototype and 0.11g for the APS-15E. In addition, the amount of 1-MG in vivo was 29.0% ± 5.8% for the APS-Prototype, significantly higher than that for the APS-15E (13.6% ± 1.9%). The desirable loss of albumin is 2–4g in hemodiafiltration, and it was 3.92 ± 1.03g for the APS-Prototype. The prototype showed excellent solute removal performance with no clinical or engineering problems.     

Molecular and cytogenetic organization of the 5S ribosomal DNA array in chicken (Gallus gallus)

The 5S ribosomal (r) RNA genes encode a small (120-bp) highly-conserved component of the large ribosomal subunit. The objective of the present research was to study the molecular and cytogenetic organization of the chicken 5S rDNA. A predominant 2.2-kb gene (5S) consisting of a coding and intergenic spacer (IGS) region was identified in ten research and commercial populations. A variant gene repeat of 0.6kb (5S) was observed in some of the populations. Genetic linkage analysis and cytogenetic localization by fluorescence in-situ hybridization assigned the 5S rDNA to chromosome 9. The 5S rDNA array was determined to be 80.2±7.0kb upon electrophoretic sizing following EcoRV digestion. Sequence analysis of 5S IGS regions revealed considerable conservation between chicken subspecies (98.4% identity) as well as homology with vertebrate Pol III promoter and regulatory sequence motifs. Minor intraindividual sequence variation within 1000bp of IGS was observed in four cloned Red Jungle Fowl (Gallus gallus gallus) 5S repeats (95.5% identity in this region). Sequence comparisons between IGS regions of 5S and 5S genes indicated two short continuous (>20bp) and many short non-continuous homologous regions as well as other conserved features such as promoter and termination motifs.     

Attenuated Lapinized Chinese Strain of Classical Swine Fever Virus: Complete Nucleotide Sequence and Character of 3′-Noncoding Region

The complete nucleotide sequence including precise 5- and 3-terminal non-coding regions (NCRs) of the attenuated lapinized Chinese strain (HCLV) of Classical Swine Fever Virus (CSFV) was determined from overlapping cDNA clones constructed by separated RT-PCR and rapid amplification of cDNA ends (RACE) methods. The genomic RNA of the HCLV strain consists of 12,310 nucleotides (nts) including 374nts and 242nts in the 5- and 3-NCRs, respectively. It contains one large open reading frame (ORF) encoding a polyprotein of 3,898 amino acids with a calculated molecular weight of 437.6kDa. There is one notable insertion of 12 continuous nts, CTTTTTTCTTTT in the 3-NCR of HCLV genomic cDNA when compared with its parental virulent Shimen strain. Sequence alignment of partial 3-NCR reveals two groups of CSFV vaccine strains carrying similar T-rich insertions at different positions in this region. Computer-predicted secondary structures suggest that T-rich insertion greatly change the structures and thus decrease the promoter functions of 3-NCRs during the replications of these two groups of CSFV vaccine strains.     

Molecular evidence supporting the classification of Hosta virus X as a distinct species of the genus

Summary. A potexvirus, Hosta virus X (HVX-Kr), causing mosaic and mottle symptoms was isolated from hosta plants (Hosta spp.) in Korea. The 3-terminal 2,711 nucleotides excluding the poly (A) tail were determined and shown to include the partial viral replicase, triple gene block (TGB) 1 (26kDa), TGB2 (13kDa), TGB3 (8kDa), and 23kDa coat protein (CP) and the 3-nontranslated region (NTR), typical of potexviruses. The CP gene of the type isolate of HVX (HVX-U) was amplified by RT-PCR and its nucleotide sequence was determined. The CPs of HVX-Kr and HVX-U had 100% and 98.9% identical amino acids and nucleotides, respectively. Most of the regions of the genome HVX had over 50% nucleotide identical to other sequenced potexviruses. This is the first report of sequence information of HVX and molecular evidence supporting the virus as a distinct species of the genus Potexvirus.Received November 29, 2002; accepted May 21, 2003 Published online July 17, 2003     

Usefulness of low-priming-volume cardiopulmonary bypass circuits and dilutional ultrafiltration in neonatal open-heart surgery

In neonate open-heart surgery, cardiopulmonary bypass (CPB) with extreme hemodilution induces an increased capillary permeability and accumulation of extravascular fluid, resulting in organ dysfunction. We evaluated the effects of a reduced priming volume for CPB and dilutional ultrafiltration (DUF) during neonatal open-heart surgery. Nineteen consecutive neonates with complete transposition of the great arteries who underwent an arterial switch operation were retrospectively assigned into two groups: the high-priming-volume circuit group (group A, n = 9) and the low-priming-volume circuit group (group B, n = 10). Patients in group B underwent surgery with a miniaturized CPB circuit and using the DUF technique. The priming volume of group B was nearly two-thirds that of group A. The water balance value after CPB and surgery was significantly lower in group B (–126 ± 118ml, –116 ± 116ml) than in group A (88 ± 218ml, 83 ± 165ml). Systolic blood pressure just after CPB was higher in group B (67.9 ± 9.1mmHg) than in group A (55.4 ± 10.3mmHg). Postoperative ventilatory support was shorter in group B (45 ± 19h) than in group A (68 ± 27h). In neonatal cardiac surgery, low-priming-volume CPB circuits and DUF improve the water balance during surgery and may attenuate any inflammatory reaction, which would help preserve postoperative organ function.     

Molecular Analysis of Fiji Disease Fijivirus Genome Segments 1 and 3

Fiji disease fijivirus (FDV) genomic segments 1 (S1) and 3 (S3) were completely sequenced. FDV S1 comprised 4532nt and was predicted to encode a 170.6kDa protein. FDV S3 comprised 3623nt and was predicted to encode a 135.5kDa protein. The terminal sequences of S1 and S3 were 5 AAGUUUUU......CAGCUAGCGUC 3 and 5 AAGUUUUU......CAGCAGAUGUC 3, respectively, and located immediately adjacent to these sequences were 12bp imperfect inverted repeats. The predicted translation product of FDV S1 showed highest similarity to Rice black-streaked dwarf virus (RBSDV) S1 and is thought to encode the viral RNA-dependent RNA polymerase (RdRp). The predicted translation product of FDV S3 was found to be most similar to RBSDV S4 which is thought to encode the 'B-spike' protein. The FDV sequence contained an ATP/GTP binding motif and a leucine zipper motif, but these motifs were not found in the RBSDV sequence. Phylogenetic analysis based on the amino acid sequences of the RdRp of FDV S1 and other reoviruses revealed that the fijiviruses form a cluster close to the oryzaviruses. The RdRp sequences were grouped into genera that were consistent with the current reovirus classification scheme that is based on physico-chemical and biological properties.     

<Emphasis Type="Italic">Rice stripe virus</Emphasis> 23.9 K protein aggregates and forms inclusion bodies in cultured insect cells and virus-infected plant cells

Summary. The genome of Rice stripe virus (RSV, genus Tenuivirus) contains seven open reading frames (ORFs). Little is known about the products of four of these ORFs, including the 23.9K protein encoded by the virus-sense ORF of RNA3. Western blotting revealed that the 23.9K protein was synthesized in the host plant and also in the planthopper vector of RSV. Using a baculovirus vector, the 23.9K protein was expressed, both unfused and fused with red-shifted green fluorescent protein, in Spodoptera frugiperda cells. Inclusion bodies were observed by light microscope in cells expressing fused or unfused proteins. Inclusion bodies in cells expressing the fused protein fluoresced under blue light. By immunoelectron microscopy, electron-dense inclusion bodies in cells expressing the unfused protein were specifically labeled with 23.9K protein antiserum. Moreover, electron-dense masses labeled with 23.9K protein antiserum were observed in virus-infected wheat tissue by electron microscopy. This paper thus demonstrates that RSV 23.9K protein can aggregate in vivo and form inclusion bodies in infected plant tissue.Received December 24, 2003; accepted June 11, 2003 Published online August 7, 2003     

Nano-scaled hydroxyapatite/polymer composite IV. Fabrication and cell adhesion properties of a three-dimensional scaffold made of composite material with a silk fibroin substrate to develop a percutaneous device

Nano-scaled sintered hydroxyapatite (HAp) particles with an a-axis length of 87 ± 23nm, a c-axis length of 236 ± 81nm, and a mean aspect ratio (c/a) of 2.72 were covalently linked onto a silk fibroin (SF) substrate chemically modified by graft polymerization with -methacryloxypropyl trimethoxysilane (MPTS). Graft polymerization with poly(MPTS) on SF was conducted by free-radical initiation in a water solvent with pentaethylene glycol dodecyl ether as a nonionic surfactant. The alkoxysilyl groups of the graft polymers avoided hydrolysis and maintained their activity in coupling with the hydroxyl groups on the HAp surface despite the use of water as the reaction solvent. The weight gain of poly(MPTS) on SF increased with increasing the reaction time, eventually reaching a plateau value of about 15wt% after 50min of reaction time. After HAp covalent coating, the particles separated or aggregated into several crystals, as shown by scanning electron microscopic observation. L929 fibroblast cells adhered more plentifully on HAp-coated SF compared to untreated SF and hydrolyzed poly(MPTS)-grafted SF during 24h or 48h of incubation. The cells adhered only on the HAp surface but not at all on the dehydrated grafted surface of SF without HAp. A button-shaped prototype for a percutaneous device was manufactured by transplantation of HAp-coated SF fibers of about 100µm in length onto silicone moldings using an adhesive, and the device showed good cell adhesiveness.     

A sandwich-ELISA for the diagnosis of Peste des petits ruminants (PPR) infection in small ruminants using anti-nucleocapsid protein monoclonal antibody

Summary. A sandwich ELISA test using PPR specific monoclonal antibody (clone 4G6) to an epitope of nucleocapsid protein has been developed. The test uses polyclonal sera to capture the antigen from clinical samples (swabs and tissues). Captured antigens from clinical samples are detected using PPR specific monoclonal antibody. The test is specific to PPR as it failed to detect rinderpest vaccine virus (RBOK strain). Varieties of clinical samples originating from laboratory experiments (n=231) and from field (n=259) were employed to test the efficacy of sandwich-ELISA test. The test compared very well with an internationally accepted commercial Immune-capture ELISA kit, which uses biotinylated monoclonal antibody against the nucleocapsid protein. On a parallel testing using 490 clinical samples, 4G6 MAb based sandwich ELISA had an overall relative diagnostic specificity of 92.8% and diagnostic sensitivity of 88.9% compared to the commercial kit. The newly developed test is free from prozone phenomenon. PPR outbreaks from various parts of India have been confirmed using the test. Findings suggested that the newly developed ELISA is suitable for PPR diagnosis under field conditions.