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Simple real-time PCR assay with one set of primer and probe for rapid, sensitive qualitative and quantitative detection of Entamoeba histolytica has been used. Consensus sequences were used to amplify a species-specific region of the 16S rRNA gene, and fluorescence resonance energy transfer hybridization probes were used for detection in a LightCycler platform (Roche). The anchor probe sequence was designed to be a perfect match for the 16S rRNA gene of Entamoeba species, while the acceptor probe sequence was designed for Entamoeba histolytica, which allowed differentiation. The performed characteristics of the real-time PCR assay were compared with ELISA antigen and microscopical detection from 77 samples of individuals with suspected clinical diagnosis of imported E. histolytica infection. Stool and liver abscess pus samples were examined with analytical sensitivity of 5 parasites per PCR reaction. The melting curve means Tms (standard deviation) in clinical isolates were 54°C. The real-time assay was 100% sensitive and specific for differentiation of Entamoeba histolytica, compared with conventional ELISA or microscopy. This real-time PCR assay with melting curve analysis is rapid, and specific for the detection and differentiation of Entamoeba histolytica. The suitability for routine use of this assay in clinical diagnostic laboratories is discussed.

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Acta Veterinaria Hungarica
Authors: Fumina Sasaoka, Jin Suzuki, Toh-Ichi Hirata, Toshihiro Ichijo, Kazuhisa Furuhama, Ryô Harasawa, and Hiroshi Satoh

. , Sato , S. and Harasawa , R. ( 2010 ): Differential detection of hemotropic Mycoplasma species in cattle by melting curve analysis of PCR products . J. Vet. Med. Sci. 72 , 77 – 79 (Erratum, J. Vet. Med. Sci. 72 , 1704

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Orvosi Hetilap
Authors: Orsolya Dóra Ács, Bálint Péterfia, Péter Hollósi, Irén Haltrich, Ágnes Sallai, Andrea Luczay, Karin Buiting, Bernhard Horsthemke, Dóra Török, András Szabó, and György Fekete

–12. 12 Wang W, Law HY, Chong SS. Detection and discrimination between deletional and non-deletional Prader–Willi and Angelman syndromes by methylation-specific PCR and quantitative melting curve analysis

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-Ortuno , M. I. , Colmenero , J. D. , Reguera , J. M. , Garcia-Ordonez , M. A. , Pachon , M. E. , Gonzalez , M. , Morata , P. : Rapid diagnosis of human brucellosis by SYBR green I-based real-time PCR assay and melting curve analysis in serum

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-mediated PCR clamping and melting curve analysis: comparison with restriction fragment length polymorphism analysis Clin Chemistry 50 481 489 . 13

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confirm the specificity of the amplification reactions, a melting curve was recorded. Each sample was replicated three times; the value of the threshold cycle (Ct) was the same as that of the corresponding mean. The relative fold expression of each mRNA

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. and Bartram, C. R. 2000: Rapid and reliable detection of Nras mutations in acute lymphoblastic leukemia by melting curve analysis using LightCycler technology. Leukemia 14 , 312-315. Rapid and reliable detection of Nras

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Shemirani AH, Muszbek L: Rapid detection of the factor XIII Val34Leu (163 G—>T) polymorphism by real-time PCR using fluorescence resonance energy transfer detection and melting curve analysis. Clin. Chem. Lab. Med. 42, 877–879 (2004

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Acta Microbiologica et Immunologica Hungarica
Authors: Károly Péter Sárvári, József Sóki, Miklós Iván, Cecilia Miszti, Krisztina Latkóczy, Szilvia Zsóka Melegh, and Edit Urbán

DNA templates. StepOne RT-PCR machine (Applied Biosystems, USA) was used for the PCR cycling and detection: 95 °C for 10 min, followed by 35 cycles of 95 °C for 15 s, 56 °C for 20 s, 72 °C for 30 s, and 1 cycle of 72 °C for 75 s and a melting curve

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Physiology International
Authors: Katsuyuki Tokinoya, Seiko Ono, Kai Aoki, Koki Yanazawa, Yasuhiro Shishikura, Takehito Sugasawa, and Kazuhiro Takekoshi

for 3 s, and annealing and elongation at 60 °C for 3 s. Melting curve analysis confirmed that the PCR product did not contain non-specific by-products. TATA-binding protein (Tbp) was used as the internal normalizing control for the mRNA. The cycle

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