Authors:Erika Orosz, Katalin Perkátai, Beatrix Kapusinszky, Ágnes Farkas, and István Kucsera
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.
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 meltingcurve analysis of PCR products . J. Vet. Med. Sci. 72 , 77 – 79 (Erratum, J. Vet. Med. Sci. 72 , 1704
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
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 meltingcurve analysis
Authors:Faham Khamesipour, Abbas Doosti, and Ebrahim Rahimi
-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 meltingcurve analysis in serum
Authors:Mahrokh Samadi, Alireza Shirpoor, Ali Taghizadeh Afshari, Fatemeh Kheradmand, Yousef Rasmi, and Maryam Sadeghzadeh
confirm the specificity of the amplification reactions, a meltingcurve 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
Authors:B. Mayr, M. Holzheu, G. Schaffner, and et al.
. and Bartram, C. R. 2000: Rapid and reliable detection of Nras mutations in acute lymphoblastic leukemia by meltingcurve analysis using LightCycler technology. Leukemia 14 , 312-315.
Rapid and reliable detection of Nras
Authors:A. Takáts, Amir-Houshang Shemirani, K. Zsóri, C. András, and Z. Csiki
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 meltingcurve analysis. Clin. Chem. Lab. Med. 42, 877–879 (2004
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 meltingcurve
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. Meltingcurve 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