Specific Hyperimmune Serum Research Articles

Immunoblot analysis of excrertory/secretory antigens of Cotylophoron cotylophorum and Gastrothylax crumenifer

The study focused on purification, characterization and isolation of excretory/secretory antigens of ovine paramphistomes. A total of 5.321 and 7.170 mg of protein was obtained from 630 adult Cotylophoron cotylophorum and 290 adults Gastrothylax crumenifer, respectively. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) (12%) analysis of purified C. cotylophrum revealed 6 polypeptides in the range of 10 to 35 kDa while a prominent band at 15 kDa with G. crumenifer. Western blot analysis of purified excretory/secretory antigens of C. cotylophorum revealed 4 immuno-determinant bands in the range of 10 to 30 kDa using their specific rabbit hyperimmune serum, whereas G. crumenifer showed 2 polypeptides between 10 to 25 kDa. The common polypeptide observed at 20 kDa in both purified excretory/secretaty antigens can be used as a candidate antigen for development of serodiagnosis in the early stage of infection.

ДИАГНОСТИКА ВОЗБУДИТЕЛЯ СОСУДИСТОГО БАКТЕРИОЗА КРЕСТОЦВЕТНЫХ МЕТОДОМ ДОТ-ИММУНОАНАЛИЗА

In the presented work, the possibility of using the dot-immunoassay method for diagnosing the causative agent of cruciferous vascular bacteriosis is considered. The disease is characterized by harmfulness, and therefore the control of the contamination of agricultural lands and crop storages with it is of great economic importance. To solve this problem, highly specific hyperimmune rabbit serum was obtained on the basis of pathogen cell wall proteins. Since the species composition of the microflora found in farmland conditions is characterized by a wide variety of genera and species, an experimental assessment was made of the ability of the proposed serum and the method as a whole to detect the pathogen under conditions close to the field. For this, extracts of white cabbage plant leaves were prepared - experimental samples taken in the field and obtained from the distribution network, and control samples grown in laboratory conditions - then checked by dot-immunoassay. As a result, it was found that the diagnostic product has specificity sufficient to detect the causative agent of cruciferous vascular bacteriosis under conditions of contamination with non-target microflora and the presence of plant tissue cells, and the chosen method is suitable for creating individual and complex rapid test systems suitable for the needs of phytosanitary control of crop production. Due to the low cost of consumables, the simplicity of working methods and the possibility of automating the process, the proposed method can be effectively implemented into everyday practice to assess the condition of the crop arriving for storage - which will help reduce its losses, improve the quality of the goods sold and reduce labor costs and working time when laying vegetables to warehouses.

Detection of lumpy skin disease virus in cattle using real-time polymerase chain reaction and serological diagnostic assays in different governorates in Egypt in 2017.

Background and Aim:Lumpy skin disease (LSD), is a highly infectious viral disease of cattle, caused by LSD virus (LSDV) which belongs to the genus Capripoxvirus of family Poxviridae. In the summer of 2017, skin lesions suggestive of LSD were observed in cattle at several governorates in Egypt. This study aimed to detect LSDV in cattle specimens using rapid serological and molecular diagnostic assays.Materials and Methods:A total of 46 skin biopsies and uncoagulated blood samples were collected from cattle with LSD suggestive clinical signs, as well as 290 coagulated whole blood samples from cattle without skin lesion in different governorates in Egypt during the summer of 2017. Skin biopsies were used for virus isolation from the chorioallantoic membrane of 11-day-old specific pathogen-free embryonated chicken eggs (SPF-ECEs). LSDV was identified using conventional polymerase chain reaction (PCR), real-time PCR (RT-PCR), and fluorescent antibody technique (FAT) with specific hyperimmune serum against LSDV. Cattle sera were examined using indirect FAT (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA).Results:Skin nodules and sitfast lesions were significant clinical signs observed in all LSD suspect cattle. SPF-ECEs, from which positive isolations were made and it showed characteristic inflammatory and focal white pock lesions. The isolated viruses were identified as LSDV by FAT, conventional gel-based PCR, and RT-PCR. Among the skin biopsies and corresponding blood samples, LSDV-positive samples percentage were 39.13 and 36.95 by RT-PCR, followed 34.78 and 28.26 by conventional PCR and then 32.6 and 26.8 by FAT, respectively. The total positive percentage of LSDV antibody detected in cattle serum samples were 17.93 and 14.48 by indirect ELISA and IFAT.Conclusion:LSDV was detected and identified in skin biopsies and corresponding blood samples of naturally infected cattle, more LSDV-positive samples were detected by RT-PCR, followed by conventional PCR and then FAT. The indirect ELISA detected more antibody-positive samples than the IFAT from cattle serum samples. The RT-PCR assay is simple, sensitive, rapid, and reliable for the detection of LSDV in blood and skin nodule biopsies of suspected cattle.

Enzymatic cleavage promotes disassembly of GII.3 norovirus virus like particles and its binding to salivary histo-blood group antigens

In this study, we found that addition of fecal extract significantly promoted the binding of GII.3 NoV VLPs to salivary HBGAs. SDS-PAGE analysis indicated that major capsid proteins (VP1) were cleaved into two major bands with molecular weights of 26 and 31kD, respectively. Pretreatment of fecal extract by boiling or addition of protease inhibitor cocktail or type specific protease inhibitor (leupeptin and aprotinin) during incubation all decreased VP1 cleavage and its binding to salivary HBGAs. Trypsin digestion led to cleavage of VP1 and promoted its binding to HBGAs, suggesting that the active enzyme(s) might be trypsin or trypsin-like enzymes. Trypsin and fecal extract pretreatment all led to loss of morphological intact VLPs, indicating enhanced signal was possible due to increased binding of fragmented subunits. N-terminal sequencing was performed to characterize the cleavage sites with indecisive results. In vitro VLP-salivary HBGAs binding blockade assay using VLPs derived from VP1 of different GII.3 strains and rabbit anti-genotype specific hyperimmune serum indicated that GII.3 NoVs might have conservative HBGA binding sites. In summary, our results provide evidence about the widespread presence of active enzyme in fecal samples that can cleave GII.3 NoV VLPs and demonstrate that GII.3 NoVs have conservative HBGA binding sites which might have implications in the design of multivalent NoV vaccines.

ДОСЛІДЖЕННЯ ТЕРАПЕВТИЧНОЇ ЕФЕКТИВНОСТІ ГІПЕРІМУННОЇ СИРОВАТКИ ПРОТИ КАЛІЦИВІРУСНОЇ ІНФЕКЦІЇ КОТІВ

In Ukraine, as elsewhere in the world, feline calicivirus is one of the most common infectious diseases of cats. The main reason is the continuous growth of infections in animal populations, and high recurrence interest of feline calicivirus in cats that have undergone treatment. In addition, nowadays, it is a chronic disease. Therefore, according to existing treatment regimens, there is a need to develop new, more effective, aimed at the elimination of the pathogen through the appointment of immunomodulators, antibiotics with simultaneous application of specific medicine, including hyperimmune serum against feline calicivirus.The article presents experimental data on the study of various schemes of complex treatment for feline calicivirus using Fosprenil, Tylozine and specific hyperimmune serum against feline calicivirus. Apart from antigenic reaction, medical hyperimmune serum show immunomodulatory properties. It is proved that the use of specific globulin at a dose of 2 ml per animal in the treatment of cats increases the effectiveness of therapy by 25% compared with the use of schemes that included only antibiotic and immune modulator. At the same time, it is known, that the use of globulin without antibiotics and immunomodulators does not show high therapeutic efficacy.

Isolation of Pigeonpox Virus from Severe Infection of Pigeons in Diyala province: Virological and Histopathological Study

Pigeonpox virus was isolated from severe cases of avipoxvirus infection affecting 64 pigeons in Ba'aquba of Diyala governarate. The virus grew well on chorioallantoic membrane of chick embryos of 11-12 days old, and produced typical pock lesions. Histopathological sections of infected tissue samples revealed typical pox lesions. The virus was identified as Pigeonpox virus using of specific hyper immune serum and indirect immuno-flourescent and indirect immuno-peroxidase tests. The virus agglutinated RBCs of pigeon, fowl, turkey and duck. Experimental infection in pigeons produced moderate infection as compared to the diseased birds, while in chicken the virus produced mild infection.

Management of an outbreak of peste des petits ruminants with antibiotic combined hyperimmune serum therapy

Peste des petits ruminants (PPR) is a highly contagious viral disease associated with high morbidity and mortality in goats of Bangladesh. Though there is no known effective drug against PPR virus, Animal Health Research Division (AHRD), Bangladesh Livestock Research Institute (BLRI) has developed a treatment technology “antibiotic combined hyperimmune serum therapy (ACHST)” against the deadly PPR virus. This article described the management of a huge outbreak of PPR by ACHST at goat farm of BLRI regional station, Naikhongchari, Bandarban, a hilly area of Bangladesh. Seventy two goats out of 159 goats were infected with PPR virus naturally. The clinical history and necropsy findings were recorded and nasal swabs were collected for diagnosis of PPR virus by RT-PCR. Thirty goats among 72 goats were died before providing treatment and the rest alive 42 goats were received ACHST. Each of the goats was treated with PPR specific hyperimmune serum @ 4-5ml/goat slow IV for once, Amcicoli-D (Ampicillin trihydrate, Colistin sulfate and Dexamethasone acetate) @ 1ml/10kg body weight daily IM once for 3 days and metronidazole @ 500mg/35kg body weight orally thrice daily for 3 days. After treatment, 38 goats (93.23%) were recovered out of 42 goats. The comparative evaluation of ACHST at different stages of naturally PPR infected goat’s shown highest recovery rate in incubation (100%) and prodromal (100%) phases followed by pneumonic (91.67%) and diarrhoeal (81.25%) phases. It may be concluded that the ACHST could be used as therapeutic interventions for PPR infected goats.Asian J. Med. Biol. Res. June 2015, 1(2): 230-234

A novel ICK peptide from the Loxosceles intermedia (brown spider) venom gland: Cloning, heterologous expression and immunological cross-reactivity approaches

The venom of a Loxosceles spider is composed of a complex mixture of biologically active components, consisting predominantly of low molecular mass molecules (3–45 kDa). Transcriptome analysis of the Loxosceles intermedia venom gland revealed ESTs with similarity to the previously described LiTx peptides. Sequences similar to the LiTx3 isoform were the most abundant, representing approximately 13.9% of all ESTs and 32% of the toxin-encoding messengers. These peptides are grouped in the ICK (Inhibitor Cystine Knot) family, which contains single chain molecules with low molecular mass (3–10 kDa). Due to their high number of cysteine residues, ICK peptides form intramolecular disulfide bridges. The aims of this study were to clone and express a novel ICK peptide isoform, as well as produce specific hyperimmune serum for immunoassays. The corresponding cDNA was amplified by PCR using specific primers containing restriction sites for the XhoI and BamHI enzymes; this PCR product was then ligated in the pET-14b vector and transformed into E. coli AD494 (DE3) cells. The peptide was expressed by IPTG induction for 4 h at 30 °C and purified by affinity chromatography with Ni-NTA resin. Hyperimmune serum to the recombinant peptide was produced in rabbits and was able to specifically recognize both the purified recombinant peptide and the native form present in the venom. Furthermore, the recombinant peptide was recognized by antisera raised against L. intermedia, L. gaucho and L. laeta whole venoms. The recombinant peptide obtained will enable future studies to characterize its biological activity, as well as investigations regarding possible biotechnological applications.

Isolation and molecular characterization of a second serotype of the encephalomyocarditis virus

Encephalomyocarditis virus (EMCV) and Theilovirus are the two species of the Cardiovirus genus. Whereas theiloviruses comprise several sero-/genotypes, all known EMCV isolates are serologically very similar and are thought to belong to one serotype, named EMCV-1. Here, a novel EMCV type is described. Strain RD 1338 (D28/05) was isolated from a wood mouse (Apodemus sylvaticus) in Germany and can be distinguished from EMCV-1 by serological and molecular means. Failure of EMCV-1 specific hyperimmune sera to neutralize RD 1338 suggests a distinct serotype. The viral genome was de novo sequenced using next-generation Illumina/Solexa technologies. Considerable differences of the BC-loop/loop I/loop II sequences of VP1, the VP2 puff and the VP3 knob provide a structural basis for deviant serological properties. Sequence alignments reveal amino acid identities of 75 percent for the P1 region and 84 percent for the P2 and P3 regions when comparing RD 1338 to EMCV-1 strains and some 60 percent and less than 50 percent amino acid identities, respectively, for comparisons with theilovirus strains. Phylogenetic analyses of the P1, 2C and 3CD gene regions support the establishment of an EMCV-2 serotype. In contrast to the theilovirus sero-/genotypes that show a narrow host range, EMCV-1 infects a wide variety of hosts. The host range of EMCV-2 remains to be determined.

Expression, purification and characterization of allelic variants of MSP-1 42 from Indian Plasmodium falciparum isolates

The C-terminal 19 and 42 kDa fragments of Plasmodium falciparum merozoite surface protein 1 (MSP-1) have shown to be protective in animals against lethal parasite challenge. The MSP-1 19 being highly conserved may lack sufficient number of T-cell epitopes in order to elicit a broader response in genetically diverse populations. The inclusion of additional epitopes from the N-terminal MSP-1 42 has shown to enhance the protective efficacy of MSP-1 19 vaccine. In an attempt to examine the strain specific immunogenicity to MSP-1, we have cloned and expressed three diverse allelic variants of MSP-1 42 from Indian P. falciparum isolates in bacteria. Among three alleles, one was extremely rare and not been found before. These purified and refolded recombinant products were recognized by conformation specific monoclonal antibodies and hyper-immune sera. Immunization of mice and rabbits with the purified proteins generated high titer biologically active polyclonal antibodies supporting further development of this vaccine candidate antigen.

ETIOLOGICAL AND MOLECULAR BIOLOGICAL INVESTIGATION OF CAPRINE HERPESVIRUS 1 ISOLATED IN BULGARIA

From goat and bucks with genital disorders and abortions the attempts for isolation of caprine herpesvirus 1 (CHV 1) were carried out. For virus exaltation Dexametazone was used. For viruses isolation vaginal, nasal, rectal, preputial swabs and organ samples were used. Primary and continuous cell cultures rabbit kidney (RK), Madin Darby bovine kidney (MDBK), and embrional bovine trachea (EBTR) were used for cultivation. For determination of DNA type and lipid envelop 60 μg/ml iod desoxiuridine (JDUR) and the ether treat ment was used. Neutralization by specific hyperimmune serum obtained from Switzerland was performed. Five CHV 1 strains were isolated by cell cultures and identified as goat herpesviruses from dif ferent Bulgarian regions. After electron microscopy the viral agents with typical size and morphology for herpesviruses were established. For demonstration gC gene of CHV 1 the polymerase chain reaction (PCR) with primers designed from sequences deposited in gene bank were developed. Isolated on cell cultures herpesviruses were proved as caprine herpesvirus 1 by using applied PCR variant. The products after gC gene amplification from Bulgarian isolates were separated on the same place as the amplicons of ref er ence CHV 1 strains.

Expression analysis of proteases of Mycobacterium leprae in human skin lesions

Proteases are commonly involved in bacterial pathogenesis and their inhibition has represented a successful therapeutic approach to treat infectious diseases. However, there is little information on the role of proteases in the pathogenesis of Mycobacteria. Five of these genes, three coding for putative secreted proteases, were selected in the present study to investigate their expression in Mycobacterium leprae isolated from skin biopsies of multibacillary leprosy patients. Via nested-PCR, it was demonstrated that mycP1 or ML0041, htrA2 or ML0176, htrA4 or ML2659, gcp or ML0379 and clpC or ML0235 are transcribed in vivo during the course of human infection. Moreover, the expression of Gcp in leprosy lesions was further confirmed by immunohistochemistry using a specific hyperimmune serum. This observation reinforces the potential role of mycobacterial proteases in the context of leprosy pathogenesis.

Cysteine proteinase 30 in clinical isolates of T. vaginalis from symptomatic and asymptomatic infected women

A cysteine proteinase of 30kDa (CP30) of Trichomonas vaginalis, is known to play a role in cytoadherence of the parasite to host cells. However, the CP30 activity in clinical isolates from symptomatic and asymptomatic patients has not been analyzed. In the present study, CP30 was detected in 20 fresh and long-term culture maintained T. vaginalis isolates each from symptomatic and asymptomatic women by substrate gel electrophoresis and immunoblotting. Though CP30 was detected in all the fresh isolates from 20 symptomatic and 20 asymptomatic women, the intensity of CP30 band was significantly higher in isolates from symptomatic as compared to asymptomatic women indicating higher expression in former. CP30 was found in all the 20 long-term cultured isolates from symptomatic whereas only in 70% of asymptomatic women indicating that CP30 expression is a more stable characteristic of symptomatic isolates. The isolates from symptomatic women, demonstrated significantly higher cytoadherence to VECs as compared to asymptomatic women. In both the types of isolates, this cytoadherence was inhibited significantly by CP30 specific hyperimmune serum. These results confirm that CP30 is an important virulence factor of T. vaginalis and has an important role in cytoadherence to VECs and thus has a role in pathogenesis of trichomoniasis.

Urinary Tract Infections due to Mycoplasma canis in Dogs

Urine samples were obtained from 100 dogs with symptoms of lower urinary tract disease by cystocentesis and were examined for mycoplasmas. Urinalysis, haematological and biochemical analyses were also performed. Bacteria were isolated from urine in 41 of 100 dogs; Mycoplasma canis was isolated from four of 100 (4%) urine samples and three were pure culture. Selective mycoplasma media were used for isolation. In growth inhibition test, propagation of the four M. canis isolates was inhibited by their specific hyperimmune sera and there was no cross reactivity between isolates and hyperimmune sera of other mycoplasmas. Dogs in which M. canis was isolated were azotemic. All dogs were treated with enrofloxacin, furosemide, and supportive therapy (fluid therapy, ascorbic acid). In all animals, clinical improvements were observed after treatment.

Identification and immunological characterization of conserved Mycoplasma hyopneumoniae lipoproteins Mhp378 and Mhp651

Mycoplasma hyopneumoniae, the etiological agent of swine enzootic pneumonia, is an important pathogen in the swine industry worldwide. Investigations on pathogenicity mechanisms as well as current serological detection methods and the development of new recombinant subunit vaccines are hampered by the lack of known and well characterized, species-specific M. hyopneumoniae antigens. As a first step to solve these problems membrane and membrane-associated proteins were enriched from M. hyopneumoniae cells by Triton X-114 fractionation and further analyzed by 2D gel electrophoresis and Western blot analyses using convalescent sera. Two previously unknown immunogenic proteins were identified by quadrupole time-of-flight mass spectrometry and database analyses as the conserved putative lipoproteins, Mhp378 and Mhp651. Both proteins were expressed as recombinant GST fusion proteins and reacted with sera from convalescent pigs. Coated as solid-phase antigen, Mhp651 showed a distinct cross-reaction only with Mycoplasma flocculare specific rabbit hyperimmune serum, whereas Mhp378 was only recognized by the positive control serum directed against M. hyopneumoniae, thereby indicating its species specificity.

Isolation of Lumpy skin disease virus form naturally infected cattle previously vaccinated with live attenuated sheep poxvirus vaccine

Lumpy skin disease virus (LSDV) was isolated, from naturally infected cattle that have a history of previous vaccination with live attenuated sheep pox virus (SPV) vaccine. The virus was isolated on chorio-allantoic membrane (CAM) of specific pathogen free (SPF) embryonated chicken eggs (ECE) and identified by agar gel precipitation test (AGPT) and neutralization test using specific hyperimmune serum against LSDV and SPV. Characteristic intracytoplasmic inclusion bodies was detected in trypsenized cell of infected CAM stained with H&E. Laboratory studies for characterization of isolated LSDV revealed that it was stable at a wide range of pH, but it was inactivated by exposure to 56 0C for 15 minutes. Treatment of isolated LSDV with lipid solvents (20% ethyle ether and chloroform) reduced the virus titer 3.2 and 4.4 log respectively after 24 hrs at 4 0C .On cross neutralization testcomplete neutralization of isolated LSDV was obtained with both reference LSDV and SPV antisera. Cattle vaccinated with live attenuated SPV vaccine under experimental condition found to be protected against natural field infection with LSDV.

Recombinant protein‐based ELISA and immuno‐cytochemical assay for the diagnosis of SARS

A new Coronavirus (SARS‐CoV) is the aetiological agent of the severe acute respiratory syndrome (SARS). Because of the critical role played by serological assays for SARS diagnosis, an in‐house ELISA based on SARS‐CoV recombinant antigens was developed. The SARS‐CoV nucleocapsid protein (N), three N fragments (N1, N2, and N3) and the intraviral domain of the membrane protein (M2) were cloned and expressed in Escherichia coli as histidine‐tagged proteins. Six reference sera from SARS patients were used to detect virus‐specific IgG in an ELISA using each recombinant protein as coating antigen. High‐titre positive reactions were detected in all SARS positive sera. The specificity of the assay appears to be high as no positive reaction was detected in the sera of 20 healthy subjects and 73 patients with non‐SARS, low‐tract respiratory infections. Specific hyper‐immune sera to SARS‐CoV and the recombinant proteins, N, N1, N2, N3, and M2 were also generated in mice and rabbits. The specificity of these sera was confirmed by an immunocytochemical assay on biochips of SARS‐CoV infected and uninfected cells. J. Med. Virol. 76:137–142, 2005. © 2005 Wiley‐Liss, Inc.

Examination of antigenic structure and some biological activities of hemagglutinin-neuraminidase (hn) and fusion (f) glycoprotein antigens of parainfluenza 3 virus, in vitro

The objective of our study was examination of the antigenic structure fusional and hemolytic activities of the surface glycoprotein HN and F antigens of purified parainfluenza (PI 3) viruses activated with 0.025 g/dl trypsin-versen (molecular weights of 112 kD, 81-82 kD and 30-31 kD), in vitro. The samples of activated PI3 virions with total protein concentrations of 0.55 and 0.27mg/ml and hemagglutinating titre of 256 and 128 HAU/0.1 ml, induced bovine turbinate(BT) cell fusion and hemolysis of guinea-pig erythrocytes. After treatment of the aforementioned samples with specific hyperimmune sera against PI 3 virus in which the hemagglutination-inhibition (HI) titre was 1:64, cell fusion was not registered. The results show that there are possibilities to use fusional and hemolytic tests for the fast detection of imunologically important glycoprotein antigens of PI 3 viruses and their identification with specific hyperimmune sera.

Modification of bacteraemia by specific antibodies and relation with mortality in a pneumococcal mouse sepsis model.

The relationship between mortality and the bacteraemic profile was investigated in a pneumococcal (serotype 6B) sepsis BALB/c mouse model where animals received protection by specific hyperimmune serum. A single intraperitoneal dose of hyperimmune serum obtained from mice immunized with the heat-inactivated strain was administered (non-diluted or diluted to 1/4 or to 1/16) to 5-mice study groups 1 h prior to intraperitoneal inoculation with the infective inoculum (3.57 x 108 cfu/ml). Blood cultures were performed daily over 15 days, with 8 microl of blood being collected from the tail vein; the samples were resuspended in Todd-Hewitt broth containing 10% trisodium citrate and plated onto blood agar for colony counting. Animals included in the control group received placebo (PBS). Mortality was 100% in control animals within the first 48 h. Hyperimmune serum decreased and delayed mortality in a dose-related trend, producing 100%, 80%, 60% and 40% survival rates at 72, 96, 144 and 360 h, with non-diluted serum. Bacteraemic profiles with maximum colony counts > or =5 x 107 cfu/ml in blood during the follow-up period were related to > or =65% probability of death, regardless of the serum dilution administered.

The association of rubella virus in congenital cataract - a hospital-based study in India.

The association of rubella virus (RV) with congenital cataract has been well established. Since the data on association of RV with congenital cataract in India are scanty, a study was done based on virus isolation from lens aspirates in patients undergoing therapeutic lensectomy and serology. To determine the incidence of the association of rubella virus with congenital cataract. The lens aspirates collected during the 9-year period (from 1990 to 1998), from 70 children up to 12 years of age with congenital cataract were processed for the isolation of rubella virus by conventional viral isolation culture method using BHK-21 and Vero cell lines. Identification of the virus was confirmed by immunofluorescence using human anti-rubella virus specific hyperimmune serum. Serum samples were collected from 55 out of these 70 children and the presence of antibodies to RV was detected by ELISA test. RV was isolated from lens aspirates in seven (10%) out of the 70 children with congenital cataract. Of the 55 sera tested, 22 had both anti-rubella IgM and IgG antibodies, in 13 only anti-RV IgG antibodies, in seven only IgM antibodies and the rest of the 13 samples did not have detectable levels of rubella antibodies. Among the children who had IgM antibodies, 12 (24.5%) were below the age of 6 months. It can be concluded based on virus isolation that 10% of patients with congenital cataract were due to rubella infection and the detection of 24.5% anti-RV IgM antibodies in children below 6 months old shows the possible association of rubella virus with congenital cataract.