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Products (7)

LightCycler^® RNA Amplification Kit HybProbe

Kit for one-step RT-PCR using the LightCycler^®Carousel-Based System.

For general laboratory use.

LightCycler^® RNA Master HybProbe

Easy-to-use reaction mix for one-step RT-PCR using the LightCycler^® Carousel-Based System.

For general laboratory use.

LightCycler^® FastStart DNA Master^PLUS HybProbe

Easy-to-use hot start reaction mix for PCR using the LightCycler^® Carousel-Based System.

For general laboratory use.

LightCycler^® DNA Master HybProbe

Easy-to-use fast high-performance qPCR kit for HybProbe signal transmission with all LightCycler^®Carousel-Based Instruments.

For general laboratory use.

LightCycler^® FastStart DNA Master HybProbe

Easy-to-use hot start reaction mix for PCR using the LightCycler^® Carousel-Based System.

For general laboratory use.

Publications (10)

Automated RNA Extraction by MagNA Pure Followed by Rapid Quantification of Cytokine and Chemokine Gene Expression with Use of Fluorescence Resonance Energy Transfer

Keywords: MagNA Pure LC ,LightCycler system ,MagNA Pure LC RNA Isolation Kit I ,Transcriptor First Strand cDNA Synthesis Kit ,LightCycler FastStart DNA Master Hybridization Probes

J. Loeffler , P. Swatoch , D. Akhawi-Araghi , H. Hebart and H. Einsele

Clin Chem , 49, 6, 955-958

Automated RNA Extraction by MagNA Pure Followed by Rapid Quantification of Cytokine and Chemokine Gene Expression with Use of Fluorescence Resonance Energy Transfer

Keywords: MagNA Pure LC ,LightCycler system ,MagNA Pure LC RNA Isolation Kit I ,Transcriptor First Strand cDNA Synthesis Kit ,LightCycler FastStart DNA Master Hybridization Probes

J. Loeffler , P. Swatoch , D. Akhawi-Araghi , H. Hebart and H. Einsele

Clin Chem , 49, 6, 955-958

Rapid quantitative detection of TE-AML1 fusion transcripts in pediatric acute lymphoblastic leukemia by real-time reverse transcription polymerase chain reaction using fluorescently labeled probes

Keywords: LightCycler DNA Master Hybridization Probes ,LightCycler FastStart DNA Master Hybridization Probes

P. Bolufer , E. Barragan , A. Verdeguer , J. Cervera , J. M. Fernandez , I. Moreno , E. Lerma , C. Esquembre , M. Tasso , V. Fuster , M. Bermudez and M. Sanz

Haematologica, 87, 1, 23 - 32

Background and Objectives: We report a new real-time reverse transcription polymerase chain reaction (RT-PCR) method for quantification of TEL-AML1 transcripts. The method is based on hybridization probe (HybProbe) chemistry applied in LightCycler equipment. The study group comprised 44 successive cases of pediatric acute lymphoblastic leukemia (P-ALL).

Design and Methods: The quantitative estimation of TEL-AML1 transcripts was performed in 10 P-ALL TEL-AML1-positive patients. The PCR was performed in capillary tubes in 10 microL final volumes using two sets of primers: M1, which detects the long (L-form) and short (S-form) transcripts, and M2, specific for the L-form. The fluorescently labeled HybProbes (TEL 3FL and TEL 5LC) hybridize to the TEL region. TEL-AML1 expression was normalized relative to the levels of AML1 transcripts. Standard curves were prepared using serial dilutions of plasmids with TEL-AML1 or AML1 inserts.

Results: The sensitivity attained allowed the detection of TEL-AML1 transcripts at a 10-4 dilution of a cDNA sample from a patient at diagnosis. The within-assay coefficient of variation (CV) for TEL-AML1 was 7.0% and the between-assay CV was 13%. Levels of TEL-AML1 transcript and the TEL-AML1/AML1 ratio decreased by more than four log units (p <0.001) during or after the course of initial treatment. Most of the patients who achieved complete remission after 5-6 months of initial treatment were TEL-AML1 negative, although some positive samples with negligible amounts of TEL-AML1 transcripts were still detected.

Interpretation and Conclusions: This method has the sensitivity and reliability required to monitor the presence of minimal residual disease, and could be a powerful tool in monitoring the efficacy of the response to chemotherapy.

Rapid quantitative detection of TE-AML1 fusion transcripts in pediatric acute lymphoblastic leukemia by real-time reverse transcription polymerase chain reaction using fluorescently labeled probes

Keywords: LightCycler DNA Master Hybridization Probes ,LightCycler FastStart DNA Master Hybridization Probes

P. Bolufer , E. Barragan , A. Verdeguer , J. Cervera , J. M. Fernandez , I. Moreno , E. Lerma , C. Esquembre , M. Tasso , V. Fuster , M. Bermudez and M. Sanz

Haematologica, 87, 1, 23 - 32

Rapid and Sensitive Detection of Shiga Toxin-Producing Escherichia coli from Nonenriched Stool Specimens by Real-time PCR in Comparison to Enzyme Immunoassay and Culture

Keywords:

T. Grys , L. Sloan , J. Rosenblat and R. Patel

J Clin Microbiol, 47, 7, 2008 - 2012

Shiga toxin (Stx)-producing Escherichia coli (STEC) bacteria are a frequent cause of food-borne gastroenteritis, hemorrhagic colitis, and hemolytic uremic syndrome. Because antimicrobial agents are generally contraindicated in patients infected with STEC, a sensitive and specific diagnostic test with rapid turnaround is essential. Current culture methods may fail to detect non-O157 STEC. We evaluated a Stx gene real-time PCR assay using hybridization probes and the LightCycler instrument with 204 prospectively collected stool specimens, which were also tested for Stx by enzyme immunoassay (EIA) (ProSpecT STEC; Remel, Lenexa, KS) and by culturing on chromogenic agar (Chromagar O157; BD BBL, Sparks, MD). In addition, 85 archived stool specimens previously tested for Stx (by EIA) and/or E. coli O157:H7 (by culture) were tested by PCR. Sample preparation for PCR included mixing the stool in sterile water and extraction of nucleic acid using the MagNA Pure LC instrument (Roche Diagnostics). The PCR assay had 100% sensitivity and specificity compared to EIA and culture for specimens collected prospectively (4 of 204 specimens were positive) and compared to culture and/or EIA for archival specimens (42 of 85 specimens were positive). Both the EIA and PCR produced positive results from a specimen containing an O103 serotype STEC in the prospective specimens, and the PCR test detected three positive specimens that contained nonviable STEC in the archived specimens. The PCR assay demonstrated 100% sensitivity and specificity compared to EIA and/or culture and more rapid turnaround than either EIA or culture.

Documents (11)

LightCycler^® Hybridization Probes

The most direct way to monitor PCR amplification for quantification and mutation detection.

(["Article"], PDF, 2373 KB) Download

PNA Clamp Technique for Detecting a Ki-ras2 Mutation Using the LightCycler Instrument

Literature indicates that point mutations in codon 12 of the Ki-ras2 gene are associated with colon cancer. The detection of a point mutation in the high background of wild-type cells is very difficult, which represents a problem for many research projects focused on processes that take place during cancerogenesis. Therefore, a quick and easy, yet reliable method of detecting single point mutations is preferred. This article shows an application example for Peptide Nucleic Acid (PNA) oligomers in combination with Hybridization Probes on the LightCycler Instrument. Here, PNAs suppress the codon 12 wild-type PCR product, as they bind to nucleic acids with higher stringency, compared to deoxyribonucleotides, and do not serve as primers for Taq polymerase. Due to these properties, PNAs can be used to detect single point mutations in research samples in a high background of wild-type sequences.

(["Article"], PDF, 282 KB) Download

Rapid Genotyping and Quantification on the LightCycler^TM with Hybridization Probes

(["Article"], PDF, 404 KB) Download

Simultaneous Detection of SNPs in Four Porcine Genes Using Hybridization Probes and the LightCycler^<® 2.0 Instrument

(["Article"], PDF, 262 KB) Download

Identification of Cytokine SNPs Using LightCycler Hybridization Probes and Melting Curve Analysis

The analysis of single nucleotide polymorphisms (SNPs) in cytokine genes has begun to shed some light on the roles of cytokines in the pathophysiology of various diseases. Allelic SNPs in the promoter region of interleukin 10 (IL-10) and tumor necrosis factor alpha (TNF-?) genes can affect the respective levels of cytokine produced.

(["Article"], PDF, 319 KB) Download

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LightCycler® Hybridization Probes

PNA Clamp Technique for Detecting a Ki-ras2 Mutation Using the LightCycler Instrument

Rapid Genotyping and Quantification on the LightCyclerTM with Hybridization Probes

Simultaneous Detection of SNPs in Four Porcine Genes Using Hybridization Probes and the LightCycler<® 2.0 Instrument

Identification of Cytokine SNPs Using LightCycler Hybridization Probes and Melting Curve Analysis

LightCycler^® Hybridization Probes

Rapid Genotyping and Quantification on the LightCycler^TM with Hybridization Probes

Simultaneous Detection of SNPs in Four Porcine Genes Using Hybridization Probes and the LightCycler^<® 2.0 Instrument