Browsing by Author "Gulhan, T."

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Comparison of Culture and Pcr for the Detection of Brucella Melitensis in Blood and Lymphoid Tissues of Serologically Positive and Negative Slaughtered Sheep
(Blackwell Publishing, 2008) Ilhan, Z.; Aksakal, A.; Ekin, I. H.; Gulhan, T.; Solmaz, H.; Erdenlig, S.
Aims: To compare the culture and PCR methods for detection of Brucella melitensis in blood and lymphoid tissue samples obtained from slaughtered sheep (n = 162) testing positive/negative in serological tests (Rose Bengal test and serum agglutination test). Methods and Results: Of 162 sheep examined, 45 were positive and 117 negative in serological tests. A PCR assay based on a pair of Br. melitensis-specific primers was used to detect DNA in blood and lymphoid tissue. Brucella melitensis was isolated from 1.2% (2/162) and 17.2% (28/162) of the blood and lymphoid tissue samples respectively. Positive PCR products with a molecular size of 731 bp were obtained from 27.7% (45/162) of blood and 29.0% (47/162) of lymphoid tissue samples. Conclusions: The species-specific PCR assay detected a higher number of Br. melitensis DNA both from serologically positive (P < 0.01 in blood PCR, P < 0.001 in tissue PCR) and serologically negative (P < 0.001 in both blood PCR and tissue PCR) sheep compared with classical bacteriological culture methods. Significance and Impact of the Study: The results emphasize the importance of using more than one type of diagnostic technique for the detection of animals positive for brucellosis, especially with epidemiological purposes.
Comparison of Pcr Assay and Bacteriological Culture Method for the Detection of Brucella Melitensis in Stomach Content Samples of Aborted Sheep Fetuses
(M H Schaper Gmbh Co Kg, 2007) Ilhan, Z.; Solmaz, H.; Aksakal, A.; Gulhan, T.; Ekin, I. H.; Boynukara, B.
The aim of this study was to evaluate the polymerase chain reaction (PCR) assay for detection of Brucella melitensis in stomach content samples of aborted sheep fetuses and to compare its performance with bacteriological culture method. It was also aimed to determine the agreement between PCR and Rose Bengal plate test (RBPT). Materials were collected from aborted sheep from 109 different sheep flocks in the region of Van during the lambing seasons of 2004-2005 and 2005-2006. Stomach contents from 135 aborted sheep fetuses were examined by bacteriological culture and PCR, and 135 sera from these aborting ewes were tested by RBPT Identification and typing of Brucella strains were performed using standard classification test. B. melitensis biovar 3 was isolated from 26 (19.2 %) of foetal stomach contents. B. melitensis was detected by PCR in 29 (21.4 %) stomach content samples. Twenty five sera (18.5 %) from aborting ewes tested positive by RBPT The detection limit of B. melitensis 16 M strain by PCR was 1.7 x 10(3) cfu (colony forming units) /ml in spiked stomach contents. Diagnostic sensitivity and specificity of the PCR were detected as 100 % and 97.2 %, respectively. The agreement between PCR and RBPT was found to be 97 %. In conclusion, PCR assay would have an advantage over conventional bacteriological culture method, but in particular for its ability to meet the specificity requirements for the detection of B. melitensis in stomach content samples of aborted sheep fetuses.
Detection of Brucella Melitensis Dna in the Milk of Sheep After Abortion by Pcr Assay
(Univ Austral Chile, Fac Ciencias veterinarias, 2008) Ilhan, Z.; Solmaz, H.; Aksakal, A.; Gulhan, T.; Ekin, I. H.; Boynukara, B.
Laboratory diagnosis of brucellosis is generally performed by microbiological and serological methods. PCR assay is a specific and sensitive choice for the detection of different bacterial agents. An evaluation of this test was carried out for the detection of Brucella melitensis DNA in sheep milk. 102 milk samples from sheep after abortion were taken and studied using bacteriological culture, PCR and milk ring test (MRT). PCR found B. melitensis DNA in 24 (23.5%) out of 102 milk samples, while only 8 (7.8%) of the samples were positive to B. melitensis through direct culture. MRT found 28 (27.4%) positive milk samples. The detection limit for PCR in sheep milk inoculated with B. melitensis strain 16 M was 1.7 x 10(3) - 1.7 x 10(4) cfu/ml. PCR and MRT coincidence was 96%. The diagnostic sensitivity and specificity were determined as 100% and 81.3% respectively for PCR assay and 75% and 75% for MRT. PCR is a useful tool for a fast diagnosis of B. melitensis in sheep milk.
Determination of Avian Influenza a Viruses in Some Avian Species in Van Lake Basin by Real Time-Pcr, Isolation and Subtyping
(2011) Boynukara, B.; Gulhan, T.; Adizel, O.; Ilhan, Z.; Aksakal, A.; Durmus, A.; Solmaz, H.
In this study, feces samples collected during 37 months from February 2006 to March 2009 from 2013 animals consisting of 47 avian species covering irregular vagrant, transit migrant, winter visitor, migratory and native birds in the Van Lake Basin Turkey were tested by Real-Time PCR (RT-PCR) with respect to Avian Influenza (AI) type A virus M2 gene. Of them, 59 samples (2.9%) were found to be positive. RT-PCR positive samples were examined with the same method with respect to H5N1 and 4 samples (6.8%) were found to be positive. RT-PCR positive 59 samples were inoculated in Embryonated Chicken Egg (ECE) and AI type A virus was isolated from 12 samples (20.3%). Of the isolates, 3 were typed as H1N7, 2 as H7N9, 2 as H11N9 and 1 as H8N4 with Hemagglutination Inhibition (HI) and Neuraminidase Inhibition (NI) tests. About 4 isolates obtained from winter visitor Anas cylpeata which had been determined as H5N1 by RT-PCR and agarose gel electrophoresis, gave positive reaction by HI test both with HI and H5 antisera and all were typed as Nl by Nl-test. Feces samples found to be positive by RT-PCR belonged to avian species Anseriformes, Charadriiformes, Pas seriformes, Gruiformes and Phoenicopteriformes orders. The highest positivity was determined in winter visitor Anas acuta (37.1%) and Anas penelope (22.5%) ducks. Of the RT-PCR positive 59 samples, 43 (72.9%) were determined in the samples collected during winter and spring of 2006-2009. Positivity was determined at a rate of 35.2% in respect of AI type A by RT-PCR in different species sharing the same time and place. With this study, the presence of AI type A viruses in various wild birds in the Van Lake Basin was determined for the first time in Turkey. © Medwell Journals, 2011.