• Title/Summary/Keyword: PCR diagnostic assay

Search Result 141, Processing Time 0.025 seconds

Caseous lymphadenitis by Corynebacterium pseudotuberculosis in a Saanen dairy goat (Capra hircus aegagrus) (Corynebacterium pseudotuberculosis에 의한 유산양의 가성결핵 증례)

  • Shin, Dong-Ho;Song, Yun-Kyung;Byun, Jae-Won;Kim, Ha-Young;Kim, Hyoung-Soon;Woo, Gye-Hyeong;Lee, O-Soo;Jung, Byeong Yeal
    • Korean Journal of Veterinary Research
    • /
    • v.50 no.1
    • /
    • pp.25-28
    • /
    • 2010
  • Caseous lymphadenitis (CLA) is a chronic and contagious disease of sheep and goats caused by Corynebacterium (C.) pseudotuberculosis. A four-year-old female Saanen dairy goat was submitted to the Animal Disease Diagnostic Center at National Veterinary Research and Quarantine Service. The clinical signs of the goat were emaciation, abortion and quadriplegia. The multifocal nodules of lymph nodes were encapsulated and filled with whitish caseous contents on the cut surface. Histopathologically, lymph nodes displayed suppurative and necrotizing granulomas. Caseous necrosis was diffusely observed in the center of the lymph nodes. Gram positive bacilli were shown in the lesions. C. pseudotuberculosis was isolated and confirmed by the biochemical tests and PCR assay. Based on clinical signs, histopathological examination and bacterial isolation, we diagnosed this case as CLA. To our knowledge, this is the first report of CLA in a Saanen dairy goat in Korea.

Serological evidence of West Nile viral infection in archived swine serum samples from Peninsular Malaysia

  • Mohammed, Mohammed Nma;Yasmin, Abd Rahaman;Noraniza, Mohd Adzahan;Ramanoon, Siti Zubaidah;Arshad, Siti Suri;Bande, Faruku;Mohammed, Hussni O.
    • Journal of Veterinary Science
    • /
    • v.22 no.3
    • /
    • pp.29.1-29.6
    • /
    • 2021
  • West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

Development and Validation of a Perfect KASP Marker for Fusarium Head Blight Resistance Gene Fhb1 in Wheat

  • Singh, Lovepreet;Anderson, James A;Chen, Jianli;Gill, Bikram S;Tiwari, Vijay K;Rawat, Nidhi
    • The Plant Pathology Journal
    • /
    • v.35 no.3
    • /
    • pp.200-207
    • /
    • 2019
  • Fusarium head blight (FHB) is a devastating wheat disease with a significant economic impact. Fhb1 is the most important large effect and stable QTL for FHB resistance. A pore-forming toxin-like (PFT) gene was recently identified as an underlying gene for Fhb1 resistance. In this study, we developed and validated a PFT-based Kompetitive allele specific PCR (KASP) marker for Fhb1. The KASP marker, PFT_KASP, was used to screen 298 diverse wheat breeding lines and cultivars. The KASP clustering results were compared with gelbased gene specific markers and the widely used linked STS marker, UMN10. Eight disagreements were found between PFT_KASP and UMN10 assays among the tested lines. Based on the genotyping and sequencing of genes in the Fhb1 region, these genotypes were found to be common with a previously characterized susceptible haplotype. Therefore, our results indicate that PFT_KASP is a perfect diagnostic marker for Fhb1 and would be a valuable tool for introgression and pyramiding of FHB resistance in wheat cultivars.

CD5+/CD21-Chronic Lymphocytic Leukemia in a Cat

  • Choi, Sorin;Bae, Hyeona;Chun, Daseul;Kim, Jihu;Shin, Sun Woo;Cho, ARom;Jung, Dong-In;Yu, DoHyeon
    • Journal of Veterinary Clinics
    • /
    • v.37 no.6
    • /
    • pp.350-354
    • /
    • 2020
  • Feline chronic lymphocytic leukemia (CLL) is a rare disease. Its diagnosis is not simple because of the absence of clinical signs and the presence of mature lymphocytosis. An 11-year-old female spayed Russian Blue cat was referred to the veterinary medical teaching hospital for lethargy, diarrhea, weight loss, and inappetence. Marked lymphocytic leukocytosis and a significantly increased number of small-to-intermediate-sized lymphocytes in the peripheral blood were found on hematological examination. The results of the feline leukemia virus and immunodeficiency virus test were negative. Further, mild splenomegaly was detected. Bone marrow aspirate analysis revealed mature lymphocytosis and a clonally rearranged T cell receptor gene with the polymerase chain reaction (PCR) for antigen receptor rearrangement assay. Flow cytometric immunophenotyping showed a homogeneous population of CD5+/CD21-T-cells in the peripheral blood and bone marrow. According to the results of the aforementioned examinations, CLL was diagnosed. Treatment was not initiated at the time of diagnosis because the clinical signs were mild and did not affect the quality of life. This report describes the clinical findings and use of advanced diagnostic tools such as molecular clonality analysis and immunophenotyping for the diagnosis of feline CLL.

Increasing of Macrophage Migration Inhibitory Factor Expression in Human Patients Infected with Virulent Brucella in Iraq

  • Khudhur, Hasan R.;Menshed, Abbas Ali;Hasan, Ahmed Abbas
    • Microbiology and Biotechnology Letters
    • /
    • v.48 no.4
    • /
    • pp.569-573
    • /
    • 2020
  • Brucellosis is a zoonotic disease caused by Brucella infections and humans usually contract this disease from close contact with infected animals or their products, usually via the ingestion of cheese or crude milk. Macrophage migration inhibitory factor (MIF) and Pro- and anti-inflammatory cytokines play an important role in susceptibility/resistance and the immunopathogenesis of Brucella infection. These cytokines are crucial factors in the initiation and progression of protective immunity against Brucella infection but the role of MIF has not been well studied in the human response to intracellular microbes. This study was designed to investigate the effect of MIF expression on Brucella susceptibility. A total of 85 positive rose Bengal tests and 24 samples from healthy individuals were collected for this study and subjected to polymerase chain reaction assays (PCR) of the bcsp31 diagnostic gene. MIF concentrations were evaluated using Enzyme-Linked immunosorbent assay (ELISA) and the results showed that 46 (54%) of the rose Bengal test samples were positive and 39 (46%) were negative for bcsp31 (p ≤ 0.05) and used as the gold standard for all of the comparisons in this study. The ELISA results indicate that the mean concentration of MIF was significantly higher in patients with positive rose Bengal tests when compared to the control groups and that its concentration increases with increasing age in both the patient and control groups (p ≤ 0.05).

Histone Deacetylases and their Inhibitors as Potential Therapeutic Drugs for cholangiocarcinoma - Cell Line findings

  • Sriraksa, Ruethairat;Limpaiboon, Temduang
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.4
    • /
    • pp.2503-2508
    • /
    • 2013
  • Histone deacetylation mediated by histone deacetylases (HDACs) has been reported as one of the epigenetic mechanisms associated with tumorigenesis. The poor responsiveness of anticancer drugs found with cholangiocarcinoma (CCA) leads to short survival rate. We aimed to investigate mRNA expression of HDACs class I and II, and the effect of HDAC inhibitors, suberoylanilide hydroxamic acid (SAHA) and valproic acid (VPA), in CCA in vitro. Expression of HDACs was studied in CCA cell lines (M213, M214 and KKU-100) and an immortal cholangiocyte (MMNK1) by semi-quantitative reverse transcription-PCR. SAHA and VPA, as well as a classical chemotherapeutic drug 5 -fluorouacil (5-FU) were used in this study. Cell proliferation was determined by sulforhodamine assay. $IC_{50}$ and $IC_{20}$ were then analyzed for each agent and cell line. Moreover, synergistic potentional of VPA or SAHA in combination with 5-FU at sub toxic does ($IC_{20}$) of each agent was also evaluated. Statistic difference of HDACs expression or cell proliferation in each experimental condition was analyzed by Student's t-test. The result demonstrated that HDACs were expressed in all studied cell types. Both SAHA and VPA inhibited cell proliferation in a dose-dependent manner. Interestingly, KKU-100 which was less senstitive to classical chemotheraoeutic 5-FU was highly was sensitive to HDAC inhibitors. Simultaneous combination of subtoxic doses of HDAC inhibitors and 5-FU signiicantly inhibited cell proliferation in CCA cell lines compared to single sgent treatment($P{\leq}0.01$), while sequentially combined treatments were less effective. The present study showed inhibitory effects of HDACIs on cell proliferation in CCA cell lines, with synergistic antitumor potential demonstrated by simultaneous combination of VPA or SAHA with 5-FU, suggesting a novel alternative therapeutic strategy in effective treatment of CCA.

White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
    • /
    • v.15 no.1
    • /
    • pp.7-13
    • /
    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

Molecular Prognostic Profile of Egyptian HCC Cases Infected with Hepatitis C Virus

  • Zekri, Abdel-Rahman N.;Hassan, Zeinab K.;Bahnassy, Abeer A.;Sherif, Ghada M.;ELdahshan, Dina;Abouelhoda, Mohamed;Ali, Ahmed;Hafez, Mohamed M.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.11
    • /
    • pp.5433-5438
    • /
    • 2012
  • Background: Hepatocellular carcinoma (HCC) is a common and aggressive malignancy. Despite of the improvements in its treatment, HCC prognosis remains poor due to its recurrence after resection. This study provides complete genetic profile for Egyptian HCC. Genome-wide analyses were performed to identify the predictive signatures. Patients and Methods: Liver tissue was collected from 31 patients with diagnosis of HCC and gene expression levels in the tumours and their adjacent non-neoplastic tissues samples were studied by analyzing changes by microarray then correlate these with the clinico-pathological parameters. Genes were validated in an independent set by qPCR. The genomic profile was associated with genetic disorders and cancer focused on gene expression, cell cycle and cell death. Molecular profile analysis revealed cell cycle progression and arrest at G2/M, but progression to mitosis; unregulated DNA damage check-points, and apoptosis. Result: Nine hundred fifty eight transcripts out of the 25,000 studied cDNAs were differentially expressed; 503 were up-regulated and 455 were down-regulated. A total of 19 pathways were up-regulated through 27 genes and 13 pathways were down-regulated through 19 genes. Thirty-seven genes showed significant differences in their expression between HCC cases with high and low Alpha Feto Protein ($AFP{\geq}600$ IU/ml). The validation for the microarray was done by real time PCR assay in which PPP3CA, ATG-5, BACE genes showed down-regulation and ABCG2, RXRA, ELOVL2, CXR3 genes showed up-regulation. cDNA microarrays showed that among the major upregulated genes in HCC are sets. Conclusion: The identified genes could provide a panel of new diagnostic and prognostic aids for HCC.

Association of Viral Infections with Risk of Human Lymphomas, Egypt

  • Kadry, Dalia Y;Khorshed, Amira M;Rashed, Reham A;Mokhtar, Nadia M
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.17 no.4
    • /
    • pp.1705-1712
    • /
    • 2016
  • Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.

Pilot Study of the Sensitivity and Specificity of the DNA Integrity Assay for Stool-based Detection of Colorectal Cancer in Malaysian Patients

  • Yehya, Ashwaq Hamid;Yusoff, Narazah Mohd;Khalid, Imran A.;Mahsin, Hakimah;Razali, Ruzzieatul Akma;Azlina, Fatimah;Mohammed, Kamil Sheikh;Ali, Syed A.
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.5
    • /
    • pp.1869-1872
    • /
    • 2012
  • Background: To assess the diagnostic potential of tumor-associated high molecular weight DNA in stool samples of 32 colorectal cancer (CRC) patients compared to 32 healthy Malaysian volunteers by means of polymerase chain reaction (PCR). Methods: Stool DNA was isolated and tumor-associated high molecular weight DNA (1.476 kb fragment including exons 6-9 of the p53 gene) was amplified using PCR and visualized on ethidium bromide-stained agarose gels. Results: Out of 32 CRC patients, 18 were positive for the presence of high molecular weight DNA as compared to none of the healthy individuals, resulting in an overall sensitivity of 56.3% with 100% specificity. Out of 32 patients, 23 had tumor on the left side and 9 on the right side, 16 and 2 being respectively positive. This showed that high molecular weight DNA was significantly (p = 0.022) more detectable in patients with left side tumor (69.6% vs 22.2%). Out of 32 patients, 22 had tumors larger than 1.0 cm, 18 of these (81.8%) being positive for long DNA as compared to not a single patient with tumor size smaller than 1.0 cm (p <0.001). Conclusion: We detected CRC-related high molecular weight p53 DNA in stool samples of CRC patients with an overall sensitivity of 56.3% with 100% specificity, with a strong tumor size dependence.