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The development of an Enzyme Linked Immunosorbent Assay for detecting Injectious laryngotrachitis viral antibodies in chicken serum Indriani, Risa; Adjid, R.M Abdul; ., Darminto; Hamid, Helmy
Indonesian Journal of Animal and Veterinary Sciences Vol 7, No 2 (2002)
Publisher : Indonesian Animal Sciences Society

The aim of this study was to develop an Enzyme-linked immunosorbent assay (ELISA) for detection of antibody against gallid herpes virus, the causal agent of infectious laryngotracheitis (ILT) in chicken. Its application in experimental chicken under laboratory condition was also evaluated. Results showed that ELISA for ILT was developed successfully with sensitivity and specificity was 98% and 97,14% respectively. The ELISA was able to determine the dynamic of antibodies respond in experimental chickens following vaccination and artificial infection with ILT virus. It was concluded that this ELISA offers a simple, sensitive and specific antibody assay for detection of antibodies against ILT virus in chickens arising from vaccination or infection. Â Key words: ELISA, antibody, chicken, Infectious laryngotrachitis

Pathogenicity and immunogenicity local isolat infectious laryngo tracheitis virus Indriani, Risa; Hamid, Helmy; Adjid, R.M Abdul; Saepulloh, Muharam
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 2 (2004)
Publisher : Indonesian Animal Sciences Society

Infectious laryngotracheitis (ILT) is an acute and contagious respiratory diseases of chicken. The virus is Gallid herpes and belong to family herpesviridae. Two local strains of ILT virus those were BGR-6 and BKS-3 were isolated and their pathogenicity and immunogenicity were further observed after five time pareses on coris allantoic of specific pathogenic free embryonated eggs. The pathogenicity of both isolates to be possible for use as seed vaccine were detected based on pathogenicity indices and antibody response. Experimental specific pathogenic free chicken in isolator cages were infected by the isolates using103EID50. ILT virus per dose. Clinical syndromes, pathological anatomic lesions, and immunological response were observed in the infected chickens and another group at uninfected chickens as a control. Results showed that either BGR-6 or BKS-3 caused clinical signs with ITPI scores of 0,05 and 0,03 respectively and there were no mortality of infected chickens. The top antibody responces of BGR-6 and BKS-3 were observed at OD 0.90 and 0.44 respectively. It can be concluded that BGR-6 and BBS-3 had low ITPI scores, but BGR-6 gave higher antibody response and can be used as a candidate for seed vaccine. Â Key words: Infectious laryngotracheitis, ILT, BGR-6, BKS-3, pathogenicity, immunogenicity

Development of a nested PCR for detection of Bovine herpesvirus-1 (BHV-1) in bovine nasal secretion and semen Saepulloh, Muharam; Adjid, R.M Abdul; T. Wibawan, I Wayan; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 13, No 2 (2008)
Publisher : Indonesian Animal Sciences Society

A nested polymerase chain reaction (nPCR) assay for detection of Bovine herpesvirus-1 (BHV-1) in bovine semen and nasal secretions was successfully developed. The nested Polymerase Chain Reaction was based on external and internal primers from the viral gD glycoprotein gene. This nPCR assay was 1000-fold more sensitive than using PCR external primer. The nested PCR has a detection limit as low as 5 ag/ml pure BHV-1 DNA and 100,75 TCID50/500 mL BHV-1 infected cells. On the other hand,Â PCR using external primer had detection limit of about 5 fg/ml pure BHV-1 DNA. Specificity studies showed that nPCR could only detect BHV-1, whereas BHV-4, PRV, PI-3 and BRSV can not be detected. In addition, nPCR was also capable in detecting BHV-1 in nasal secretion samples from animal without clinical signs. A total of 405 samples consisted of 381 nasal secretion and 24 fresh semen samples have been tested with the nPCR. The result revealed that from 381 nasal secretion samples tested, 14 samples showed to be positive (3.68%), consisting of 13 out of 294 (4.42%) nasal secretion samples collected from Pangalengan West Java, and 1 out of 87 (1.54%) samples collected from Bogor. Furthermore, 2 out of 11 (18.18%) extended semen samples collected from Bogor and 2 out of 13 (15.38%) fresh semen samples collected from Pasuruan also showed to be positive. In addition, the nPCR was faster and easier to perform than the standard viral isolation test. It is concluded that, the nPCR can be used as test of choice for routine diagnosis of BHV-1. Key Words: Nested PCR, BHV-1, Semen, Glycoprotein D Gene, TCID50

Pathogenicity and immunogenicity local isolat infectious laryngo tracheitis virus Risa Indriani; Helmy Hamid; R.M Abdul Adjid; Muharam Saepulloh
Jurnal Ilmu Ternak dan Veteriner Vol 9, No 2 (2004): JUNE 2004
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Infectious laryngotracheitis (ILT) is an acute and contagious respiratory diseases of chicken. The virus is Gallid herpes and belong to family herpesviridae. Two local strains of ILT virus those were BGR-6 and BKS-3 were isolated and their pathogenicity and immunogenicity were further observed after five time pareses on coris allantoic of specific pathogenic free embryonated eggs. The pathogenicity of both isolates to be possible for use as seed vaccine were detected based on pathogenicity indices and antibody response. Experimental specific pathogenic free chicken in isolator cages were infected by the isolates using103EID50. ILT virus per dose. Clinical syndromes, pathological anatomic lesions, and immunological response were observed in the infected chickens and another group at uninfected chickens as a control. Results showed that either BGR-6 or BKS-3 caused clinical signs with ITPI scores of 0,05 and 0,03 respectively and there were no mortality of infected chickens. The top antibody responces of BGR-6 and BKS-3 were observed at OD 0.90 and 0.44 respectively. It can be concluded that BGR-6 and BBS-3 had low ITPI scores, but BGR-6 gave higher antibody response and can be used as a candidate for seed vaccine. Key words: Infectious laryngotracheitis, ILT, BGR-6, BKS-3, pathogenicity, immunogenicity

The development of an Enzyme Linked Immunosorbent Assay for detecting Injectious laryngotrachitis viral antibodies in chicken serum Risa Indriani; R.M Abdul Adjid; Darminto .; Helmy Hamid
Jurnal Ilmu Ternak dan Veteriner Vol 7, No 2 (2002): JUNE 2002
Publisher : Indonesian Center for Animal Research and Development (ICARD)

The aim of this study was to develop an Enzyme-linked immunosorbent assay (ELISA) for detection of antibody against gallid herpes virus, the causal agent of infectious laryngotracheitis (ILT) in chicken. Its application in experimental chicken under laboratory condition was also evaluated. Results showed that ELISA for ILT was developed successfully with sensitivity and specificity was 98% and 97,14% respectively. The ELISA was able to determine the dynamic of antibodies respond in experimental chickens following vaccination and artificial infection with ILT virus. It was concluded that this ELISA offers a simple, sensitive and specific antibody assay for detection of antibodies against ILT virus in chickens arising from vaccination or infection. Key words: ELISA, antibody, chicken, Infectious laryngotrachitis

Development of a nested PCR for detection of Bovine herpesvirus-1 (BHV-1) in bovine nasal secretion and semen Muharam Saepulloh; R.M Abdul Adjid; I Wayan T. Wibawan; Darminto .
Jurnal Ilmu Ternak dan Veteriner Vol 13, No 2 (2008): JUNE 2008
Publisher : Indonesian Center for Animal Research and Development (ICARD)

A nested polymerase chain reaction (nPCR) assay for detection of Bovine herpesvirus-1 (BHV-1) in bovine semen and nasal secretions was successfully developed. The nested Polymerase Chain Reaction was based on external and internal primers from the viral gD glycoprotein gene. This nPCR assay was 1000-fold more sensitive than using PCR external primer. The nested PCR has a detection limit as low as 5 ag/ml pure BHV-1 DNA and 100,75 TCID50/500 mL BHV-1 infected cells. On the other hand, PCR using external primer had detection limit of about 5 fg/ml pure BHV-1 DNA. Specificity studies showed that nPCR could only detect BHV-1, whereas BHV-4, PRV, PI-3 and BRSV can not be detected. In addition, nPCR was also capable in detecting BHV-1 in nasal secretion samples from animal without clinical signs. A total of 405 samples consisted of 381 nasal secretion and 24 fresh semen samples have been tested with the nPCR. The result revealed that from 381 nasal secretion samples tested, 14 samples showed to be positive (3.68%), consisting of 13 out of 294 (4.42%) nasal secretion samples collected from Pangalengan West Java, and 1 out of 87 (1.54%) samples collected from Bogor. Furthermore, 2 out of 11 (18.18%) extended semen samples collected from Bogor and 2 out of 13 (15.38%) fresh semen samples collected from Pasuruan also showed to be positive. In addition, the nPCR was faster and easier to perform than the standard viral isolation test. It is concluded that, the nPCR can be used as test of choice for routine diagnosis of BHV-1. Key Words: Nested PCR, BHV-1, Semen, Glycoprotein D Gene, TCID50

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