Double Immunodiffusion Test Research Articles

CASE REPORT: SEVERE JUVENILE TYPE PARACOCCIDIOIDOMYCOSIS WITH HEPATITIS C

We present the case of a 34-year-old man with previous history of hepatitis C and severe juvenile type of paracoccidioidomycosis characterized by involvement of the reticuloendothelial system, eosinophilia, lung compromise (pleural effusion), retinal hemorrhage, and blood culture positive to Paracoccidioides brasiliensis. The immune evaluation showed four precipitation lines to P. brasiliensis by the double immunodiffusion test. Treatment with amphotericin B and itraconazol resulted in resolution of the disease.

Role of RLIP76 in lung cancer doxorubicin resistance: I. The ATPase activity of RLIP76 correlates with doxorubicin and 4-hydroxynonenal resistance in lung cancer cells

RLIP76 functions as an ATP-dependent transporter of amphiphilic chemotherapeutic drugs such as doxorubicin (DOX, adriamycin), as well as of glutathione-conjugates of endogenous electrophilic toxins such as 4-hydroxynonenal (4HNE). RLIP76 couples transport and ATP-hydrolysis with a 1:1 stoichiometry, making the ATPase activity of RLIP76 an excellent surrogate for its transport activity. Present studies were performed to determine the relationship of the RLIP76 ATPase activity with DOX and 4HNE resistance in a panel of 13 native human lung cancer cell lines. RLIP76 was purified from each cell line and homogeneity demonstrated by SDS-PAGE and amino acid composition analysis. Anti-RLIP76 antibodies were shown by Ouchterlony double immunodiffusion tests to be non-cross-reactive with any other proteins including P-glycoprotein (Pgp) or multidrug resistance associated protein (MRP). These antibodies completely immunoprecipitated ATPase activity of purified RLIP76 fractions, further confirming homogeneity of purified RLIP76. RLIP76 ATPase purified from NSCLC cell lines was about 2-fold more active than that from SCLC in the absence of the stimulator dinitrophenyl S-glutathione (206+/-47, n=7 vs. 94+/-22, n=6, nmol/min/mg protein, respectively), or in its presence (340+/-60, n=7 vs. 186+/-32, n=6, nmol/min/mg; p<0.01). Partial tryptic digest revealed a 44 kDa internal fragment of RLIP76 beginning at Thr-294 in NSCLC cell lines. This fragment was absent from all SCLC, suggesting the possibility that the activity of RLIP76 in SCLC and NSCLC is differentially regulated through post-translational modifications. Taken together, these findings suggest that RLIP76 activity is a general determinant of 4HNE and DOX resistance, and that its activity contributes to the drug-resistant phenotype of NSCLC.

Suppressive Effect of Honey on Antigen/Mitogen Stimulated Murine T Cell Proliferation

It has been reported that the intraperitoneal administration of honey suppressed the induction of antigen-specific humoral antibody response in mice as determined by passive cutaneous anaphylaxis and Ouchterlony double immunodiffusion tests. Hence, we studied the effect of honey on induction of antigen-specific IgG antibody response and found suppression in mice as determined by ELISA and also T cell proliferation in vitro. T cell proliferation induced by antigens such as ovalbumin, diphtheria toxoid and tetanus toxoid, as well as the mitogens concanavalin A and phytohemagglutinin, was significantly suppressed by various doses of natural and commercial honeys, as evaluated by 3 H-thymidine incorporation. Furthermore, the suppressive effect of honey was also confirmed on antigen- or mitogen-induced T cell proliferative responses using splenic T cells from different haplotypes of mice. The results obtained in this work confirm the antiproliferative activity of honey.

Synthesis of novel sialic acid-containing polymers as inhibitors of hemagglutination

Influenza viruses are serious pathogens for humans. They are known to infect host tissues by first binding to sialoconjugates on the cell surface through their hemagglutinin. α-Sialoside-containing polymers have been demonstrated to be potent inhibitors of the hemagglutination on human erythrocytes by influenza viruses. Moreover, they have showed superior antigenic properties than their corresponding natural sialoconjugates from the basis of multivalent and cluster effects. For the viewpoint of drug targeting, we have tried to develop efficient synthesis of novel sialopolymers in which the inter-sialoside distances could be controlled. We report the novel synthesis of sialocopolymers using condensation polymerization. The condensation of branched α-sialoside, prepared from p-nitrophenyl α-sialoside and 3,3′-iminobis(propylamine) with 1,4-diisocyanatobutane and 1,6-diisocyanatohexane, produced copolymers without any promoters. These copolymers retained their specific lectin-binding properties, as determined with wheat germ agglutinin (WGA), a plant lectin known to bind sialosides and glucosaminides on the agar gel double immunodiffusion test. Each copolymer showed clear binding. On micro-turbidimetric assay, they exhibited strong cross-linking properties with WGA, and copolymer including the hexyl group showed stronger binding properties than the butyl one.

봉독의 항독소(IgY)생산을 위한 실험적 연구

This study was carried out for production of neutral antibody to bee venom . Hen layings were injected repeatedly with bee venom and phospholipase with Freund`s adjuvant. Specific antibody in egg yolk from immunized hen laying was separated, and purified, also immunological characteristics of anti phospholipase was invested. The results were summarized as follows: 1. Phospholipase was showed single band at molecular weight 17,000 in SDS-PAGE and bee venom was showed two band at molecular weight 17,000 and under molecular weight 6,500 in SDS-PAGE. 2. During 70 days after hen immunized with bee venom and phospholipase , antibodies(anti-bee venom IgY) to bee venom were showed poor ELISA value in egg yolk, but antibodies to phospholipase in egg yolk were increased ELISA value from 8 days or 15 days and found maximum ELISA value at 42 days. Also after booster at 49 days, ELISA value of anti Phospholipase in egg yolk was supported at optical density(O.D) 1.0 level, continuously. 3. Titer of phospholipase was showed 1: 32,000. 4. In double immunodiffusion test to phospholipase after double dilution of anti-phospholipase , only precipitation line was made in 1:1 dilution well of anti-Phospholipase . But In immunodiffusion test to anti-phospholipase after double dilution of phospholipase , Precipitation line to 250ul/ml well of phospholipase was showed. In double immunodiffusion test to bee venom(1mg/ml) after double dilution anti-phospholipase , all well without 1:32 dilution well were showed strong precipitation line. 5. In dot bloting test to anti-phospholipase after diluting bee venom(0.5mg/ml), dot bloting color was showed clearly to in bee venom.

DEVELOPMENT OF AN ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR AGGLUTININ, AN ANTIBACTERIAL PROTEIN OF THE SHRIMP FENNEROPENAEUS INDICUS

Antibacterial activity of Fenneropenaeus indicus hemolymph was mediated by Agglutinin (Ag), a protein of molecular weight 181 kDa with two subunits 97 kDa and 84 kDa. Agglutinin was purified by gel filtration chromatography, PAGE, and SDS-PAGE, and polyclonal antibodies (anti-Ag) were raised in rabbits. In a double immunodiffusion test, single precipitin lines were found between the center well containing immune serum and surrounding wells containing purified Ag and crude hemolymph. The IgG fraction was purified from anti-Ag immune serum and used to develop ELISA. The Ag standard curve was linear over a range of 17.8–8,900 ng. The sensitivity of the ELISA was 178 ng/ml. Agglutinin-free hemolymph yielded background optical density values, confirming the specificity of the assay. Serial dilutions of agglutinin-rich hemolymph produced sample titration curves that are parallel to the Ag standard curve. Thus, the ELISA is suitable for quantification of Ag in hemolymph.

Characterization and RNA2 nucleotide sequence of broad bean wilt virus 2 isolate P158

Broad bean wilt virus 2 (BBWV2) isolate P158 was characterized and the complete nucleotide sequence of viral genomic RNA2 was determined. P158 has 30 nm diameter isometric particles. Double immunodiffusion test suggested that P158 has high antigenicity homology with BBWV2 isolate B935. SDS-PAGE result showed that the coat protein of P158 was comprised of two types of polypeptide with molecular weight of 44.7 kD and 21.9 kD, respectively. The genome of P158 was made up of two RNA molecules with the length of 6.0 kb and 3.6 kb, respectively. cDNA of RNA2 was cloned by RT-PCR and sequenced. RNA2 was composed of 3597 nucleotide (nt) residues excluding the poly(A) tail and contained single long open reading frame extending from nt 230 to nt 3424 in the viral sense RNA, and encoded a polyprotein ofMr 119002 (119K). Comparison of the polyprotein and the counterpart of isolate B935 indicated that the polyprotein was cleaved at Q/G (465/466) and Q/A (867/868), to release three mature proteins: a protein of unknown function, large and small subunit. Sequence comparisons of P158 with fabaviruses showed P158 had very high sequence homology with BBWV2 isolates and patchouli mild mosaic virus, but to a less extent with BBWV1 isolates.

Development of an Enzyme-Linked Immunosorbent Assay (Elisa) for Agglutinin, an Antibacterial Protein of the Shrimp Fenneropenaeus Indicus

Abstract Antibacterial activity of Fenneropenaeus indicus hemolymph was mediated by Agglutinin (Ag), a protein of molecular weight 181 kDa with two subunits 97 kDa and 84 kDa. Agglutinin was purified by gel filtration chromatography, PAGE, and SDS-PAGE, and polyclonal antibodies (anti-Ag) were raised in rabbits. In a double immunodiffusion test, single precipitin lines were found between the center well containing immune serum and surrounding wells containing purified Ag and crude hemolymph. The IgG fraction was purified from anti-Ag immune serum and used to develop ELISA. The Ag standard curve was linear over a range of 17.8–8,900 ng. The sensitivity of the ELISA was 178 ng/ml. Agglutinin-free hemolymph yielded background optical density values, confirming the specificity of the assay. Serial dilutions of agglutinin-rich hemolymph produced sample titration curves that are parallel to the Ag standard curve. Thus, the ELISA is suitable for quantification of Ag in hemolymph.

Isolation, purification, and characterization of pregnancy-specific protein B from elk and moose placenta.

Pregnancy-specific protein B (PSPB) was isolated, purified, and partially characterized from elk and moose placenta. The procedure, which was monitored by bovine PSPB (bPSPB) RIA, included homogenization and extraction in aqueous solution, acidic and ammonium sulfate precipitation, and ion exchange, gel filtration, and affinity chromatographies. The estimated molecular sizes of moose PSPB (mPSPB) were 58 kDa and 31 kDa, and of elk PSPB (ePSPB) were 57 kDa, 45 kDa, and 31 kDa by SDS-PAGE. The isoelectric points of mPSPB were 4.8, 6.6, and 6.7, and of ePSPB were 4.8, 4.9, 6.1, and 6.2 as determined by isoelectric focusing and two-dimensional gel electrophoresis. The carbohydrate contents of mPSPB and ePSPB were approximately 3.15% and 4.98%, respectively. Although ePSPB and mPSPB were recognized by anti-bPSPB in an Ouchterlony double immunodiffusion test, they were found to share identical epitopes and partial identities compared to bPSPB. After treatment at different temperatures (20-60 degrees C) for 1 h, the immunoreactivities of ePSPB and mPSPB in serum were very stable. Only ePSPB in serum treated at 60 degrees C lost some immunoreactivity. After alteration of serum pH (pH 3-11) for 2 h, the immunoreactivities of ePSPB and mPSPB became lower at pH 3 and 4, and remained stable from pH 5 to 11. These data show that moose and elk PSPB have properties similar to those of bovine and ovine PSPB.

Cross-antigenicity Between Ruminal and Hepatic Paramphistomes and Liver Flukes of Buffalo Origin

Maji, B.P., Dwivedi, P., Rao, J.R. and Yadav, S.C. 1999. Cross-antigenicity between ruminal and hepatic paramphistomes and liver flukes of buffalo origin. J. Appl. Anim. Res., 16: 53–57. The crossantigencity among paramphistomes, Paramphistomum epiclitum, Gastrothylax crumenifer, Gigantocotyle explanatum and the common liver fluke Fasciola gigantica of buffalo was studied by Ouchterlony double immunodiffusion (DID) test. When soluble whole worm antigens of paramphistomes was probed against parasite specific rabbit hyperimmune sera, marked cross-antigenicity was in evidence not only among paramphistomes, but also with the common liver fluke of buffaloes. Two to three cross-reactive antigens were common among paramphistomes while Fasciola gigahtica showed a distinct common antigen with the three predominant paramphistomes of buffalo origin.

A Newly Established ELISA Showing the Effect of the Androgenic Gland on Secondary-Vitellogenic-Specific Protein in the Hemolymph of the Crayfish Cherax quadricarinatus

A quantitative enzyme-linked immunosorbent assay (ELISA) was developed to monitor the onset of secondary vitellogenesis in Cherax quadricarinatus females and in intersex individuals (having both male and female reproductive systems) after removal of the androgenic gland (AG). As a prerequisite for the assay, the 106-kDa polypeptide was separated from newly laid C. quadricarinatus eggs by SDS–PAGE, and anti-106-kDa antibody was raised in rabbit. The specificity of the anti-106-kDa polypeptide for proteins specific for the hemolymph of secondary-vitellogenic females was confirmed by double immunodiffusion and immunoblot cross-reactivity tests. A characteristic standard ELISA curve, using egg high-density lipoprotein (HDL), showed linearity between 16 and 500 ng (r=0.953) and was sensitive for amounts as low as 8 ng. The inter- and intraassay coefficients of variance were 14.8 and 7.2%, respectively. Only traces of egg HDL equivalents were detected in the hemolymph of male and primary-vitellogenic females (11 to 110 μg/ml), confirming the specificity of the assay, whereas high levels of such a protein (8–35 mg/ml) were detected in the hemolymph of secondary-vitellogenic females. Removal of the AG from intersex individuals leads to a significant increase in the concentration of vitellogenic-specific protein in the hemolymph (up to 2 mg/ml). Moreover, a significantly lower concentration was found in females subjected to AG transplant (79.3 μg/ml). The ELISA thus provided an accurate and sensitive tool to investigate the influence of the AG on the expression of a vitellogenic-specific protein in female and intersex C. quadricarinatus, confirming the central role of this gland in tuning sexual plasticity in this species.

Fine binding specificities to Ascaris suum and Ascaris lumbricoides antigens of the sera from patients of probable visceral larva migrans due to Ascaris suum

Fine binding specificities to Ascaris suum and A. lumbricoides antigens of the sera from patients with probable visceral larva migrans (VLM) due to A. suum infection were examined. Although multiple-dot enzyme-linked immunosorbent assay (ELISA) was found to be useful for the primary screening of patients, identification of the responsible species was sometimes difficult due to extensive cross reactions with other ascarid parasite antigens. Fine resolution to determine the causative pathogen was obtained by a rather classical Ouchterlony's double immunodiffusion test. The difference in the binding of the patients' sera to A. suum and A. lumbricoides antigens was also demonstrated by an inhibition ELISA. The patients' antibodies bound with higher avidity to the A. suum antigen than to the A. lumbricoides and Toxocara canis antigens. Combination of at least two different immunological assay methods is recommended for the diagnosis of VLM due to ascarid parasites.

Clinical significances of antibodies to Ro/SS-A autoantigens and its subtypes in primary Sjögren's syndrome.

ObjectiveTo evaluate the patterns of Ro autoantigen recognition in Korean patients with primary Sjogren’s syndrome (SS) and to investigate its clinical significance in SS.MethodsSera from primary SS (n=51) and systemic lupus erythematosus (SLE) (n=132) were tested by double immunodiffusion test and immunoblotting for reactivity with 60kDa and 52kDa Ro/SS-A proteins. Clinical manifestations were evaluated on the basis of the presence of anti-Ro/SS-A antibodies and anti-60 kDa/52kDa proteins.ResultsThe prevalence of anti-Ro/SS-A antibodies in Korean patients with primary SS was 64.7%. In immunoblotting analysis, the incidence of anti-60kDa without anti-52kDa was lower in patients with SS(3.0% vs. 11.6%, p>0.05), whereas anti-52kDa without anti-60kDa was more common in SS patients than in SLE patients(42.5% vs. 4.3%, p<0.001). Patients with anti-Ro/SS-A antibody were significantly associated with the presence of vasculitis, hyperglobulinemia and rheumatoid factor in primary SS (p<0.05).ConclusionThe patterns of 52kDa and 60kDa Ro autoantigen recognition were quite different in the SLE and primary SS. Anti-52kDa without anti-60kDa antibody may be used as a diagnostic marker for primary SS. Although the presence of anti-Ro/SS-A antibody was closely associated with certain clinical features in SS, these clinical manifestations were not correlated with the presence of antibodies against each 52kDa and 60kDa proteins. Extended studies with a large population are required to determine the clinical correlation of autoantibodies against each peptides or epitopes of Ro/SS-A proteins.

Detection of Both Subgroups I and II of Cucumber Mosaic Cucumovirus and Their Satellite RNAs on Pepper in Argentina.

Pepper plants (Capsicum annuum L.) showing stunting, mild to severe mosaic, leaf narrowing and deformation, prominent veins, fruit atrophy or malformation, and necrotic patterns on leaves and fruits were collected from commercial and experimental farms from seven districts in the central western neighboring provinces of Mendoza and San Juan. Cucumber mosaic cucumovirus (CMV) was identified directly from field samples from crude RNA extracts by reverse transcription-polymerase chain reaction (RT-PCR) (1). The PCR-amplified products were analyzed by restriction enzyme digestion with PvuII and VspI to distinguish the CMV subgroups. Serological assays (double immunodiffusion tests and indirect triple antibody sandwich enzyme-linked immunosorbent assay) with subgroup-specific monoclonal and polyclonal antisera also differentiated between the subgroups, although with lower sensitivity than found with PCR. Both subgroups I and II were found, no mixed infections were detected, and subgroup membership was related to geographical distribution: subgroup-I samples were detected in San Juan sites (north), and samples collected from the southern area (Mendoza) were of subgroup II. Some samples were sap-transmitted and passed repeatedly through tobacco and pepper. Satellite RNAs (satRNAs) were detected from total RNA extracts by RT-PCR with specific primers (5'-end oligonucleotide: 5'GGGTTATATCTGCGTGAG3' and 5'CACGGAGATCAGCATAGC3' as 3'-end primer). One satRNA isolate from each area was amplified and cloned. Three and five clones, respectively, were sequenced, obtaining 313/315 nucleotide and 334 nucleotide consensus sequences that appear to be similar (98.2 and 98.9%, respectively) to CMV-Y satRNA. This is the first report of CMV subgrouping and detection of CMV satRNAs in Argentina. Reference: (1) H. Rizos et al. J. Gen. Virol. 73:2099, 1992.

Evaluation of various diagnostic techniques for Trypanosoma evansi infections in naturally infected camels

One hundred and eight camels ( Camelus dromedarius) from Trypanosoma evansi endemic areas of the Thar Desert of Rajasthan State, India, were evaluated by various diagnostic tests including parasitological tests (wet blood film — WBF, stained thick blood film), chemical test (mercuric chloride), biological test (mouse subinoculation — MSI), and immunodiagnostic tests based on antibody detection (double immunodiffusion test — DID, card agglutination test — CATT), antigen detection (double antibody sandwich enzyme linked immunosorbent assay — Ag-ELISA). Of the tested camels 49 were found infected using the WBF of which nine gave false negative results with the mercuric chloride test. The efficacy of MSI was 87.03 percent, while the mercuric chloride test was 60.18 percent efficient. The diagnostic efficacy of CATT (72.22 percent) was found to be much better than DID (28.70 percent). Ag-ELISA was 86.11 percent efficient in detecting trypanosomal antigens. A good correlation was found between the positive results obtained by wet blood film, CATT and Ag-ELISA. It was inferred that CATT can be used to study the seroprevalenee of T. evansi with great ease, however, trypanosome antigen detection may give a more accurate idea of the prevalence of T. evansi in an endemic area.

Lectin biosynthesis in callus culture established from seeds ofCanavalia virosa

Plant tissue cultures established from isolated embryos or cotyledons were used to investigate the lectin biosynthesis inCanavalia virosa. The lectin (CVL) was detected by double immunodiffusion and hemagglutination tests. CVL was present in all callus cultures. The stem and leaf of the plantlet generated from the embryo also contain CVL, but no CVL was detected in the roots. As compared to mature seeds, callus derived from cotyledon of immature seeds exhibited the largest CVL content.

Immunohistochemical and immunocytochemical localization of .ALPHA.2u-globulin in the kidneys of female rats.

We investigated the presence of α2u-globulin, which has generally been thought to be a protein specific to male rats, in the kidneys of female rats. Immunohistochemical, immunoelectron microscopic and immunochemical studies were performed on the kidneys of male and female F344 rats using rabbit anti-serum prepared with α2u-globulin purified from male rat urine. In male rats, hyaline droplets containing granules positive for immunohistochemical staining of α2u-globulin were observed in the proximal convoluted tubular epithelium. In females, fine granules positive for staining of α2u-globulin were observed in the same region, although so-called hyaline droplets were not observed in HE-stained specimens. Upon immunoelectron microscopic examination, gold particles indicating the presence of α2u-globulin were found localized in the lysosomes of proximal convoluted tubular epithelial cells in both sexes. Furthermore, immunohistochemistry revealed that α2u-globulin was localized in the submaxillary glands in both sexes and in the liver in males. In the immunochemical study, a double immunodiffusion test, precipitin lines were formed between the anti-serum and the kidney preparation and between the anti-serum and the submaxillary gland preparation from female rats. Spur formation was observed between the precipitin lines for the anti-serum and the submaxillary gland preparation and the precipitin lines for the anti-serum and urine in female rats; therefore, a partial difference in the antigenicity of α2u-globulin between males and females was suggested. From these results, we concluded that α2u-globulin exists in the kidneys in the female rats, and it is considered to originate mainly from the submaxillary glands, not the liver.

Ein Vergleich von vier Immunoblot-Systemen zur Differenzierung von Antikörpern gegen nukleäre und zytoplasmatische Antigene

Since recently, immunoblots for detection and differentiation of antibodies against nuclear antigens (ENA), other nuclear antigens and cytoplasmic antigens, blotted on nitrocellulose strips (Western blot technique) became commercially available. The aim of our analysis was to compare the specifity and sensitivity of four different immunblot systems (blots A, B, L, V). We examined 20 sera of patients with various clinically confirmed connective tissue diseases. The sera contained a total of 32 different antigens. The antibodies in the sera were defined by double immunodiffusion (Ouchterlony) test, immunofluorescence (HEp2 cells) and by comparison with WHO reference sera.

Characterization of the main plasma lipoproteins from the ovoviviparous viperid snake Bothrops jararaca

Plasma from estrogen treated males of Bothropsjararaca fractionated by ultracentrifugation showed three main fractions with very low density, intermediate density and high density, respectively. The very low density and high density fractions are mainly composed of lipoproteins detected by specific staining with Sudan Black. Electron microscopy of the very low density lipoproteins shows a single population of particles with a mean diameter of 29.6 nm similar to that of very low density lipoproteins from laying hens. The very low density lipoprotein of Bothrops jararaca is composed of two polypeptides with molecular masses of 450 and 20 kDa. Three main polypeptides are detected in the plasma high density fraction with molecular masses of 190, 160 and 110 kDa. Analysis of whole plasma proteins, from control and estrogen treated males, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the 20 kDa very low density lipoprotein polypeptide and the 160 and 110 kDa high density lipoprotein polypeptides are induced de novo after estrogen treatment. These results suggest that the two very low density lipoprotein polypeptides are the snake counterparts of chicken very low density polypepüdes. The estrogen induced high density lipoprotein polypeptides probably make up the vitellogenin(s) of Bothropsjararaca. An antiserum raised against the whole very low density plasma fraction was able to recognize only the two polypepüdes found in this fraction in the plasma of treated males. Using this anüserum, in a double immunodiffusion test, we were able to detect only a faint reaction against Bothrops jararaca egg yolk proteins.

Isolation and Partial Characterization of Peanut Top Paralysis Virus (PTPV), a Destructive Virus Causing a New Disease in Peanut (Arachis hypogaea L.)

AbstractA destructive virus, causing top paralysis to peanut, was discovered in the wild germplasm collection growing in the USDA‐ARS greenhouses, Stillwater, Oklahoma, USA. The symptoms observed on the wild plant were restricted to a few leaves as green batches in a light green to yellow background with some leaflets having lost most of the basal part of the laminae leaving the top portion rolling upwards forming a cone. The virus was mechanically transmitted to cultivated peanut (Arachis hypogaeaL,.) where it caused more severe and destructive symptoms including stunting, severe malformation of leaves and partial or complete disappearance of leaflet laminae. This virus differed in symptomology, host range, and/or serological reactivity from allpeanut viruses reported in the literature, particularly those causing leaf malformation and stunting. The virus induced necrotic local lesions on Phaseolus vulgaris L. cv. “Topcrop” and chlorotic local lesions with necrotic centres bordered withvery bright intense red color on Chenopodium amaranticolor. In both passive indirect enzyme‐linked immunosorbent assay (PAS‐ELISA) and Ouchterlony double immunodiffusion test, the virus did not react with antisera against brome mosaic, bean yellow mosaic, peanut stripe, potato Y, tobacco mosaic, watermelon mosaic 1, watermelon mosaic 2, wheat soilborne mosaic, wheat streak mosaic, and zucchini yellow mosaic viruses.However, in reciprocal cross reactions the virus seemed to share a common antigenic determinant with a peanut mottle virus isolate from Oklahoma (PMV‐OK). The virus had flexuous filamentous particles with a length of 750–850 nm, falling within the range reported for the potyvirus group. The virus was successfully purified and the molecular weight of its protein subunit was found to be 30000 d. A polyclonal antiserum was raised in rabbits against the virus and used for reciprocal serological tests.