Thaha Rasool

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I am working in the field of Animal Genetics and Biotechnology for the last 40 years. Currently working on molecular genetics of racing camel

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Differentiation of Sheep Pox and Goat Poxviruses by Sequence Analysis and PCR-RFLP of P32 Gene

Virus Genes, 2004

Sheep pox and Goat pox are highly contagious viral diseases of small ruminants. These diseases we... more Sheep pox and Goat pox are highly contagious viral diseases of small ruminants. These diseases were earlier thought to be caused by a single species of virus, as they are serologically indistinguishable. P32, one of the major immunogenic genes of Capripoxvirus, was isolated and sequenced from two Indian isolates of goat poxvirus (GPV) and a vaccine strain of sheep poxvirus (SPV). The sequences were compared with other P32 sequences of capripoxviruses available in the database. Sequence analysis revealed that sheep pox and goat poxviruses share 97.5 and 94.7% homology at nucleotide and amino acid level, respectively. A major difference between them is the presence of an additional aspartic acid at 55th position of P32 of sheep poxvirus that is absent in both goat poxvirus and lumpy skin disease virus. Further, six unique neutral nucleotide substitutions were observed at positions 77, 275, 403, 552, 867 and 964 in the sequence of goat poxvirus, which can be taken as GPV signature residues. Similar unique nucleotide signatures could be identified in SPV and LSDV sequences also. Phylogenetic analysis showed that members of the Capripoxvirus could be delineated into three distinct clusters of GPV, SPV and LSDV based on the P32 genomic sequence. Using this information, a PCR-RFLP method has been developed for unequivocal genomic differentiation of SPV and GPV.

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Genotype differentiating RT-PCR and sandwich ELISA: Handy tools in epidemiological investigation of foot and mouth disease

Journal of Virological Methods, 2007

Years of molecular epidemiological surveillance has revealed co-circulation of two antigenically ... more Years of molecular epidemiological surveillance has revealed co-circulation of two antigenically divergent genotypes of foot and mouth disease virus serotype A in India. Genotype differentiating RT-PCR and sandwich ELISA were developed as fast, cost-effective and user-friendly alternatives to 1D region based phylogeny for detection and differentiation of genotype VI and VII. The RT-PCR assay targeting 1D region was found to be more sensitive and authentic in distinguishing genotypes than sandwich ELISA. These assays promise to be reliable tools in the epidemiological investigation of foot and mouth disease in the country.

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Development of an Indirect Elisa for Detection of E. COLI Antibodies in Cow Serum using A Recombinant Ompt as Antigen

Journal of Immunoassay and Immunochemistry, 2014

Animal Husbandry Practices at High Altitude (> 6000 Feet) In Kumaon Region of Uttrakhand, India

… Research for Rural …, 2007

Survey indicates that farmers in the study area are totally dependent on the locally available fe... more

One-step Multiplex RT-PCR Assay for the Detection of Peste des petits ruminants Virus in Clinical Samples

Veterinary Research Communications, 2006

A single-tube one-step multiplex RT-PCR was standardized to amplify both 337 bp and 191 bp fragme... more A single-tube one-step multiplex RT-PCR was standardized to amplify both 337 bp and 191 bp fragments of N and M genes of peste des petits ruminants virus (PPRV), respectively, and only a 337 bp fragment of N gene of Rinderpest virus (RPV). The RT-PCR using purified viral RNA was easily adopted for direct detection of PPRV in clinical field samples and its differentiation from RPV. The amplified N and M gene products were confirmed to be PPRV-and RPV-specific by their size in 1.5% agarose gel and restriction analysis. In the assay, the Qiagen one-step RT-PCR kit containing the Ominiscript and Sensiscript reverse transcriptases and Hot star Taq DNA polymerase was utilized. The sensitivity of the assay was found to be 100 fg of PPRV RNA. Compared with a two-step assay, the one-step assay is easier and time-saving as it requires just a single buffer for both reactions, reverse transcription (RT) and PCR. In experimentally infected goats, PPRV was detectable by the one-step RT-PCR in nasal and ocular swabs 7-17 days post infection (p.i.). and in oral swabs 7-15 days p.i. Out of 32 clinical field samples tested, 18 were positive by sandwich ELISA (S-ELISA), while 22 were positive by the one-step RT-PCR.

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Detection of Infectious bursal disease virus by ELISA using an antipeptide antibody raised against VP3 region

Acta virologica, 2004

Antigenic determinant analysis was carried out on VP3, one of major immunogenic proteins of Infec... more

Molecular cloning and expression profile analysis of interleukin-10 and interleukin-18 cDNA of Indian water buffalo (< i> Bubalus bubalis</i>)

The cDNAs encoding the interleukin-10 and interleukin-18 of Indian water buffalo (Bubalus bubalis... more The cDNAs encoding the interleukin-10 and interleukin-18 of Indian water buffalo (Bubalus bubalis) were cloned and sequenced. A 537 bp IL-10 cDNA fragment and a 623 bp IL-18 cDNA fragment were amplified by reverse transcriptase polymerase chain reaction (RT-PCR) from concanavalin A stimulated splenocytes. Sequence analysis of these cytokines revealed high level conservation at nucleic acid and protein level. Both these cytokines also showed strict conservation in the predicted secondary structure and critical amino acid residues compared to the ruminant homologues. Basal level expression of both IL-10 and IL-18 was observed in liver, lung and spleen. The expression level of IL-10 was not affected by mitogenic stimulation, whereas IL-18 was up regulated upon stimulation. The availability of these cytokine molecules will aid in the study of their role in the immunology and pathogenesis of infections in water buffalo.

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Nucleic Acid and Immunological Diagnostics for SARS-CoV-2: Processes, Platforms and Pitfalls

Accurate diagnosis at an early stage of infection is essential for the successful management of a... more Accurate diagnosis at an early stage of infection is essential for the successful management of any contagious disease. The COVID-19, caused by the SARS-CoV-2 virus is a pandemic that has affected 214 countries affecting more than 30.8 million people causing 0.957 million deaths as of third week of September, 2020. The primary diagnosis of the infection is done either by the molecular technique of RT-qPCR by detecting portions of the RNA of the viral genome or through immunodiagnostic tests by detecting the viral proteins or the antibodies produced by the host. As the demand for the test increased rapidly many naive manufacturers entered the market with novel kits and more and more laboratories also entered the diagnostic arena making the test result more error-prone. There are serious debates globally and regionally on the sensitivity and specificity of these tests and about the overall accuracy and reliability of the tests for decision making on control strategies. The significanc...

Role of selected mutations in exon 28 and 39 ofMyosin15 gene in autosomal recessive nonsyndromic sensorineural deafness among affected South Indian families

Amer J Med Genet, 2003

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Milk constitutes in non descriptive buffaloes reared at high altitudes of Kumaon hill of central Himalaya

Buffalo Bulletin

Animal use as draft and transport purpose at high altitude in Kumaon hill of Uttaranchal

Seasonal nematode egg output in goats at high altitude of Uttaranchal, India

Efficacy of avermectins against gastrointestinal nematodes of Dairy cattle in temperate region of Uttaranchal

Livestock husbandry scenario at high altitude Kumaon Himalaya

The livestock husbandry scenario during 2004-06 in some selected villages of Nainital district (U... more The livestock husbandry scenario during 2004-06 in some selected villages of Nainital district (Uttarakhand) was assessd from a total of 300 respondents. The information about the existing housing, breeding, milking, feeding and health care practices of livestock was documented. Most of the farmers in the study area kept their animals in kachha house having poor lighting and drainage facility. Breeding of animals is mainly through natural service with available bulls and the artificial insemination (AI) service was at the primitive level (21%). The milking practice involved mostly stripping method and the farmers were very much (80%) aware of clean milk production and they sale their milk to dairy co-operatives. The livestocks were mostly dependent on the locally available feeding resources like oak leaves and unclassified grasses in the village grazing land/forest area for feeding of their livestock round the year. A very few farmers were aware of vaccination programme against impo...

Identification of interleukin-26 in the dromedary camel (Camelus dromedarius): Evidence of alternative splicing and isolation of novel splice variants

Molecular Immunology, 2015

Interleukin-26 (IL-26) is a member of the IL-10 family of cytokines. Though conserved across vert... more Interleukin-26 (IL-26) is a member of the IL-10 family of cytokines. Though conserved across vertebrates, the IL-26 gene is functionally inactivated in a few mammals like rat, mouse and horse. We report here the identification, isolation and cloning of the cDNA of IL-26 from the dromedary camel. The camel cDNA contains a 516 bp open reading frame encoding a 171 amino acid precursor protein, including a 21 amino acid signal peptide. Sequence analysis revealed high similarity with other mammalian IL-26 homologs and the conservation of IL-10 cytokine family domain structure including key amino acid residues. We also report the identification and cloning of four novel transcript variants produced by alternative splicing at the Exon 3-Exon 4 regions of the gene. Three of the alternative splice variants had premature termination codons and are predicted to code for truncated proteins. The transcript variant 4 (Tv4) having an insertion of an extra 120 bp nucleotides in the ORF was predicted to encode a full length protein product with 40 extra amino acid residues. The mRNA transcripts of all the variants were identified in lymph node, where as fewer variants were observed in other tissues like blood, liver and kidney. The expression of Tv2 and Tv3 were found to be up regulated in mitogen induced camel peripheral blood mononuclear cells. IL-26-Tv2 expression was also induced in camel fibroblast cells infected with Camel pox virus in-vitro. The identification of the transcript variants of IL-26 from the dromedary camel is the first report of alternative splicing for IL-26 in a species in which the gene has not been inactivated.

Molecular characterization and expression of caprine (Capra hircus) interleukin-18 cDNA

International Journal of Immunogenetics, 2005

A Vero cell-attenuated Goatpox virus provides protection against virulent virus challenge

Acta virologica, 2004

An Indian isolate of Goatpox virus (GTPV) was adapted and propagated in Vero cells for developmen... more An Indian isolate of Goatpox virus (GTPV) was adapted and propagated in Vero cells for development of an attenuated virus. The virus was initially passaged in primary lamb testes cells and subsequently in Vero cells. At the 55th passage, the virus showed evidence of attenuation when tested for safety in seronegative goats. At this stage, the virus was found to be completely non-pathogenic. The virus was passaged further and the 60th passage was used for testing its immunogenicity in goats. The latter were inoculated with 10, 100 and 1000 TCID50 of the attenuated virus by intradermal (i.d.) route and challenged after 28 days with virulent GTPV. The attenuated virus produced no adverse reaction even at the highest dose and conferred complete protection even at the lowest dose against challenge with a high dose (2 x 10(6) of 50% skin-reactive dose SRD50) of virulent virus. Increased levels of virus-specific serum antibodies could be demonstrated by both indirect enzyme-linked immunosor...

Cloning and sequencing of truncated gIV glycoprotein gene of an Indian isolate of bovine herpesvirus 1

Acta virologica, 1999

Bovine herpesvirus 1 (BHV-1) has been reported from the Indian subcontinent in late 70's. In ... more Bovine herpesvirus 1 (BHV-1) has been reported from the Indian subcontinent in late 70's. In order to identify the origin of an Indian isolate of BHV-1 (IBR/H 167 VS) and its molecular relationship to known strains of BHV-1, a 680 bp region of the glycoprotein gene gIV was amplified by polymerase chain reaction (PCR), cloned and sequenced. Comparison of this sequences with the corresponding one of an European strain of BHV-1 (Cooper) revealed more than 99% nucleotide (nt) homology. We conclude that the Indian isolate under study does not differ from the Cooper strain regarding the gIV gene.

Nucleotide sequence of DR-B exon of major histocompatibility complex from an Indian zebu cattle breed

Indian journal of experimental biology, 1998

The Indian cattle is known to be more tolerant to tropical infections than the European cattle. I... more The Indian cattle is known to be more tolerant to tropical infections than the European cattle. In order to verify the genetic basis of this variation, the DR B exon-2 of the major histocompatibility locus, known for coding the antigen recognition site, from the Hallikar breed of Indian cattle was amplified by PCR, cloned and sequenced. Comparison of this sequence with the information available on taurus cattle brought out six unique nucleotide changes and three amino acid changes. The amino acid positions were at 17, 72 and 87. A major variable region was observed at amino acid position 85 to 87 from all the alleles so far reported for the bovine locus.

High frequency of connexin26 (GJB2) mutations associated with nonsyndromic hearing loss in the population of Kerala, India

International Journal of Pediatric Otorhinolaryngology, 2009

Objective: Mutations in connexin 26 gene (GJB2) are the most common cause of hearing loss in diff... more Objective: Mutations in connexin 26 gene (GJB2) are the most common cause of hearing loss in different populations. The aim of our study was to determine the prevalence of GJB2 mutations in the population of Kerala, India. Methods: This study was conducted on the genomic DNA of 86 affected subjects and their relatives from 59 families of Kerala, India. Mutation detection was done by sequencing and PCR-RFLP. Results: 36% of the probands had mutations in the GJB2 gene. We found that 45% (15/ 33) of the families that had a family history of deafness had mutations in GJB2 gene. Two different mutations were identified. W24X mutation was detected in 32.5% of the affected patients. Analysis of control samples revealed a carrier frequency of 0.0357 for this mutation. The estimation of haplotype frequency revealed that there was a significant association between the W24X mutation and the haplotype in this region with respect to the markers, D13S143 and D13S175 suggesting a founder effect for this mutation in this population. A novel mutation, R32L was detected in 3.5% of the affected patients. Structural prediction revealed that this mutation alters the helical structure of the first transmembrane domain of GJB2 protein resulting in defective gap junctions. Conclusion: Mutations in connexin26 is responsible for 36% of non-syndromic sensorineural deafness in the population of Kerala, India.

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