![]() ![]() Finally, the sum of positive values obtained by both DNA tests (dot blot assay plus PCR) increased the rate of positivity for ATL patients to 76.4%. A comparative study between dot blot and PCR assays performed with the blood of mice that had been experimentally infected with tachyzoites gave similar results: 60 and 70% positive results, respectively. gondii DNA detection for 10 patients (58.8%). PCR assays were performed in parallel for patients with ATL, resulting in T. We found positive hybridization signals for 12 (66.7%) of 18 patients with confirmed CT, 9 (52.9%) of 17 patients with ATL, and 2 (66.7%) of 3 TRs. We studied a total of 84 individuals: 38 patients and 46 controls. We applied a dot blot hybridization assay to blood samples for the diagnosis of cerebral toxoplasmosis (CT), acute toxoplasmic lymphadenopathy (ATL), and disseminated toxoplasmosis in transplant recipients (TRs). Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis.We report the potential use of a specific Toxoplasma gondii DNA probe (ABGTg7). Smith GR, Clarke ML, de Velde RV, Dale JL (1994) Chemiluminescent detection of Fiji disease virus with biotinylated DNA probes. Cold Spring Harbor Laboratory Press, New York Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Roy BP, Abou Haidar MG, Alexander A (1989) Biotinylated probes for the detection of potato spindle tuber viroid (PSTV) in plants. Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specificity by nick translation with DNA polymerase I. Owens RA, Diener TO (1981) Sensitive and rapid diagnosis of potato spindle tuber viroid disease by nucleic acid hybridization. Maule AJ, Hull R, Donson J (1983) The application of spot hybridization to the detection of DNA and RNA viruses in plant tissues. Gillespie D, Spiegelman S (1965) A quantitative assay for DNA/RNA hybrids with DNA immobilized on a membrane. Garger SJ, Turpen T, Carrington JC, Morris TJ, Jordan RL, Dodds JA, Grill U (1983) Rapid detection of plant RNA viruses by dot-blot hybridization. Science 161:529–540įeinberg AP, Vogelstein B (1983) A technique of radiolabeling DNA restriction fragments to high specific activity. Proc Natl Acad Sci U S A 77:6851–6855īritten RJ, Kohne DE (1968) Repeated sequences in DNA. Ann Appl Biol 127:321–328īrandsma J, Miller B (1980) Nucleic acid spot hybridization: rapid quantitative screening of lymphoid cell lines for Epstein-Barr viral RNA. Proc Natl Acad Sci U S A 74:5350–5354īlok VC, Ziegler A, Scott K, Dangora DB, Robinson DJ, Murant AF (1995) Detection of groundnut rosette umbravirus infections with radioactive and non-radioactive probes to its satellite RNA. Key wordsĪlwine JC, Kemp DJ, Stark GR (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl paper and hybridization with DNA probes. If non-radioactive labels are used, then suitable detection methods through either chromogenic or chemiluminescence is used. If radioactive probes are used, then the detection is done through autoradiography. The labels can be either radioactive or non-radioactive. The RNA probes are labelled by in vitro transcription. The DNA probes may be labelled using different methods, namely nick translation, random primed labelling or by polymerase chain reaction. The process involves isolation of nucleic acid from test plant it’s spotting on a membrane, pre-hybridization, hybridization using labelled probe and their detection. The double-stranded hybrid molecule is then detected using appropriate method depending on the reporter molecule used. The labelled probe is then allowed to form hybrid with nucleic acid isolated from the test plant. ![]() In this assay, a probe which is a single-stranded nucleic acid (either DNA or RNA) is ‘labelled’ with a reporter molecule. It is based on the homology between two strands of nucleic acid (DNA:DNA, DNA:RNA or RNA:RNA). The dot-blot hybridization is a nucleic acid hybridization technique where complementary single-stranded sequences of the probe (either RNA or DNA) hybridizes with single-stranded sequences of the test samples (either RNA or DNA) under suitable conditions of temperature and salt concentration. ![]()
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