The foundationfocus cdxbrca assay is intended to be performed with serial numbcontrolled er instruments. The qiagen hc2 test is a sandwich capture molecular hybridization assay. Comparison with hybrid capture i and pcr results in cervical lesions. Hybrid capture assay, version hc2 digene, gaithersburg, md, usa which is able to detect. Targeted enrichment by solutionbased hybrid capture to. Comparison of onclarity human papillomavirus hpv assay. Evaluation of polymerase chain reaction pcr method and. Rlu values are captured by assayspecific software on a dedicated computer attached to the luminometer.
The bdna method was initially developed by chiron emeryville, ca, usa and marketed by bayer diagnostics emeryville, ca, usa, and the hybrid capture method was developed and marketed by digene corporation gaithersburg, md, usa. Hybrid capture assay the hybrid capture assay was performed according to the directions of the manufacturer. The hybrid capture 2 hc2 system hpv dna test by digene by brenda palacios objectives state the 2 main lowrisk hpv dna types state the 2 main highrisk hpv dna types state what type of test method is used for the detection of hpv state the minimum sample volume required for testing state what the molecular sandwich consist of human papillomavirus hpv primary. Successful hybrid capturebased next generation sequencing ngs is influenced by a range of parameters including probe design efficiency, ontarget coverage, uniformity of coverage, analytical sensitivity and specificity, library complexity, amount of dna input required, scalability, reproducibility, ease of use, and overall cost.
The pdf24 creator brings you a lot more tools to create pdf files. The performance characteristics of hcii assay and the pcr for detection of hpv dna have been compared in cervical lesions and results shown variation. Comparing the cervista hpv hr test and hybrid capture 2. This genomic technique, also called exome sequencing or whole exome sequencing was first applied by using an arraybased hybrid capture method in 2007 hodges et al. The assistant opens, where you can save or choose to do something else with the new file. The assay is using s1 nuclease, which degrades singlestranded dna and rna into oligo or mononucleotides, leaving intact doublestranded dna and rna. Hybrid capture definition of hybrid capture by medical. Hybridcapture 2 assay has been the recommended method by the fda committee as a gold standard for diagnosis of hpv in clinical samples. Molecular diagnostics of human papillomavirus laboratory. Hpv genotype detection using hybrid capture sample. In hybridcapture methods, the formation of probelibrary fragment dna duplexes are used to recover regions of interest. Clinical relevance of the borderline results of the hybrid. Pdf detection of human papillomavirus dna by the hybrid. The analytical performance for in vitro rna and cultured cells was satisfactory.
Chapter 7 methods for detection of hpv infection and its clinical. The digene highrisk hpv dna test using hybrid capture. Detection of human papillomavirus dna by the hybrid capture assay article pdf available in brazilian journal of infectious diseases 72. Hybrid capture article about hybrid capture by the free. Get a printable copy pdf file of the complete article 680k, or click on a page. Qiagen, gmbh, hilden, germany, which detects dna of high. To improve the limited analytical accuracy of hc2 test results near the cutoff value 1.
Nov 27, 2018 targetenrichment strategy using hybrid capture was originally developed for human genomic studies for which it was used to capture and sequence the entire human exome. Such data will permit objective interpretation of this new procedure, and could contribute to cancer prevention. Proprietary blockers for dual and singleindex adapters, to reduce nonspecific adapter interaction during probe hybridization and increase ontarget capture performance. The original hybrid capture assay hc1 was approved by the fda in 1995. Using the illumina hiseqtm 4000 platform, hybridcapture selected libraries are sequenced to uniform depth targeting 500x coverage with 99% of exons at coverage 100x. The assay is called hybrid capture hc whole genome amplification wgaluminex lmx and is described as follows. Abstractthe diagnostic performance of the widelyused cervista hpv hr test was compared to the hybrid capture 2 hc2. The pdf24 creator then takes the screen capture and creates a pdf file. This assay uses a nucleic acid hybridisation assay where the samples with the target dna. The nuclease hybridization assay, also called s1 nuclease cutting assay, is a nuclease protection assay based hybridization elisa. For simulating a mutation at a specific position, start and end bed coordinates should denote that exact same position. Screen capture to pdf capture your screen as a pdf file. Hpv dna testing by the hcii assay method was performed with the automated hcii assay system located in the department of pathology, universiti sains malaysia, according to the protocol of the manufacturer. An in vitro nucleic acid hybridization assay with signal amplification using microplate chemiluminescence for the qualitative detection of human papillomavirus hpv types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68 in cervical specimens.
Dec 10, 2018 in hybridcapture methods, the formation of probelibrary fragment dna duplexes are used to recover regions of interest. The hybrid capture 2 hc2 system hpv dna test by digene by brenda palacios objectives state the 2 main lowrisk hpv dna types state the 2 main highrisk hpv dna types state what type of test method is used for the detection of hpv state the minimum sample volume required for testing state what the molecular sandwich consist of human papillomavirus hpv primary etiological agent in cervical. The foundationfocus cdxbrca assay uses a customdeveloped analysis pipeline to identify brca12 base substitutions and short insertiondeletions indels up to bp. Comparative study between the hybrid capture ii test and. While wes has proven useful for diagnosis and management in specific clinical situations, selected gene panels have more utility at lower cost. Hybrid capture assay definition of hybrid capture assay by. For highrisk hpv only the swabs were adequate using one of the laboratory tests digene hybrid capture ii, but both the tampon and swab were acceptable when a second test roche reverse line blot assay was used. En hybrid capture system rotary shaker 1 user manual qiagen. Clinical validation in the context of a hybrid capturebased. Commonly used signal amplification technologies include branched dna bdna and hybrid capture hc assays. The capture assay was designed to capture 600 kb of target sequence which corresponded to 1. Sep 24, 2019 the hybrid capture 2 hc2 highrisk hpv dna assay serves as a triage test in the slovenian national cervical cancer screening programme zora. First year medical students from philadelphila college of osteopathic medicine georgia campus d.
The hpv tests included the hybrid capture tube test hct, the microplatebased hybrid capture ii test hc ii, and the my09my11 l1 consensus primer pcrbased assay. Dec, 2014 first year medical students from philadelphila college of osteopathic medicine georgia campus d. This document details the lab protocol for nhanes 2003. Criteria for clinical reporting of variants from a broad.
Human papillomavirus detection by the hybrid capture 2. The chosen targeted hybrid capture sequencing assay was further developed by adding target genes of interest figure 2a in accordance with the evolving literature. Amplificationbased methods have excellent specificity and uniformity. Comparing the cervista hpv hr test and hybrid capture 2 assay. Human papillomavirus testing with the hybrid capture 2 assay. What is the process for capturing rna or dna cells lysed, rnases inactivated, attached to tube are multiple targetspecific probes, sample rna binds to these capture probes, amplification occurs as several sets of probes bind to target andor each other. A branched dna assay begins with a dish or some other solid support e.
Sample size and no longitudinal arm limited the clinical assessment of assay. Hybrid capturebased human papillomavirus detection versus pcr. Multicenter comparison of the digene hybrid capture cmv dna assay version 2. Optimizing the hybrid capture ii human papillomavirus test t. The performance characteristics of pcr and the hc2 assay for testing for hpv have been compared, and the results have varied 2, 10, 14, 19, 24, 26.
This process is experimental and the keywords may be updated as the learning algorithm improves. Currently, human papillomavirus hpv dna tests validated in large trials and epidemiological studies are the hybrid capture secondgeneration hc2 hpv dna assay and a variety of polymerase chain reaction pcr protocols. Hybrid capture is a signal amplification technique, meaning that the chemiluminescent or fluorescent signal is amplified to aid detection, rather than the target dna being amplified by pcr. Lab medicine a proprietary system used to detect and monitor viraleg, chlamydia spp, cmv, hbv, hpv infections.
Hybrid capture assay definition of hybrid capture assay. Clinical validation in the context of a hybrid capture. In addition, printed copies of each assay, together with quality control calculations, are stored in notebooks. Chr 8 point mutation indel a a a a a a a a a a a chr 10 hybrid capturebased ngs technology parallel detection of point mutations, indels, copy number changes and gene fusions reliable analysis also on small amounts of material. Highlights this hpv mrna hybridcapture assay has use in hpv transcription studies. Briefly, hc2 is a signal amplification assay that uses a combination of antibody capture.
Hybrid capture 2 hc2 highrisk hpv dna test digene catalog number 51011296 for in vitro diagnostic use a nucleic acid hybridization microplate assay with signal amplification for the chemiluminescent detection of human papillomavirus hpv types 16,18,31,33,35,39,45,51,52,56,58,59 and 68 in cervical specimens reagents for 96 tests. The flexible, fully customizable design accommodates up to 15 mb of. Hybrid capture ii, a new sensitive test for human papillomavirus. Hybrid capture ii residual swabs were kept at 4c in thinprep test bottles. Class of 2018 get together to utilize active learning an. The aptima hpv assay versus the hybrid capture 2 test in. Human papillomavirus testing with the hybrid capture 2. These values are downloaded to an excel file for data storage and subsequent file transport to westat. Thednarnahybridsarecapturedon amicroplate,andtheemittedlightismeasuredinaluminometerasrela. Principle of hybrid capture 2 hc2 technology qiagen. The samples were analyzed for the presence of highriskhpv. Hybridcapture based sequencing assays to detect novel.
Comparative study between the hybrid capture ii test and pcr. Frontiers hybrid capturebased next generation sequencing. The digene hc2 hpv dna test using hc2 technology is a signal amplified hybridization antibody capture assay that. En hybrid capture system rotary shaker 1 user manual. Briefly, hc2 is a signal amplification assay that uses a combination of antibody capture and chemiluminescent signal detection. The assay is called hybrid capture hcwhole genome amplification wgaluminex lmx and is described as follows. Part 2 of msc molecular diagnostics lecture on hybrid capture ii technology and hpv detection. Assessment of capture and ampliconbased approaches for. First, there is a sequencespecific sample enrichment process to isolate singlestranded hpv dnas while removing much of the nonspecific genomic dna that may interfere with some assays. Targetenrichment strategy using hybrid capture was originally developed for human genomic studies for which it was used to capture and sequence the entire human exome.
Pdf evaluation of the digene hybrid capture ii assay. This document details the lab protocol for nhanes 20032004. Free fulltext pdf articles from hundreds of disciplines, all in one place crossreactivity profiles of hybrid capture ii, cobas, and aptima human papillomavirus assays. Dec 01, 2016 part 2 of msc molecular diagnostics lecture on hybrid capture ii technology and hpv detection. Recentaly, a secondgeneration assay with improved diagnostic sensitivity has been developed known as hybrid capture ii test hcii and approved by the us food and drug administration fda. The hybrid capture 2 hc2 highrisk hpv dna assay serves as a triage test in the slovenian national cervical cancer screening programme zora.
Crossreactivity profiles of hybrid capture ii, cobas, and. Human papillomavirus detection by the hybrid capture 2 hc2. The only procedure that may be capable of recognizing all. A secondgeneration commercial hpv test, the hybrid capture 2 hc2 assay, was introduced as a readytouse test for routine diagnostics and includes negative controls and positive controls pcs. To implement the larger 2400 gene panel, we utilized a different target.
The hybrid capture assay is a standardized assay that is simple to perform and can utilize whole blood specimens that have been stored for up to 48 h. Technical information summary all other reagents, materials and equipment needed to perform the assay are used exclusively in the foundation medicine laboratory. Print bookmark share pdf 1880kb english format file size language download get adobe reader. Optimized buffers, cot dna, and streptavidin beads for use with xgen lockdown probes and panels in hybrid capture target enrichment. Nextera tagmentation coupled with optimized target capture ensures the fastest enrichment workflow time for custom content. Usefulness of hybrid capture hpv dna assay as a diagnostic.
Specifically, a sixperson working group of hematopathologists, clinicianoncologists, and translational scientists c. The original hybrid capture assay hc1 was approved by. Hybridcapture article about hybridcapture by the free. Sep 23, 2010 recentaly, a secondgeneration assay with improved diagnostic sensitivity has been developed known as hybrid capture ii test hcii and approved by the us food and drug administration fda. Optimizing the hybrid capture ii human papillomavirus test. Cellular heterogeneity affects the composite mrna ratio of samples. Assessment of capture and ampliconbased approaches for the. Nextera rapid capture custom enrichment is an allinone assay for library preparation and custom target enrichment. Hybrid capture 2 hc2 technology serves as the platform for qiagens nucleic acid hybridization assay for detection of human papillomavirus hpv. Evaluation of the hybrid capture assay for determination.
Evaluation of the hybrid capture assay for determination of. The hc2 highrisk hpv dna test using hybrid capture2 hc2 technology is a nucleic acid hybridization assay with signal amplification using microplate chemiluminescence for the qualitative detection of thirteen highrisk types of human papillomavirus hpv dna in cervical specimens. In the sandwich hybridization elisa assay format, the antigen ligand and antibodies in elisa are replaced with a nucleic acid analyte, complementary oligonucleotide capture and detection probes generally, in the case of nucleic acid hybridization, monovalent salt concentration and temperature are controlled for hybridization and wash stringency, contrary to a traditional. Briefly, in a micro tube, rna probes were hybridized with hpv dna which had been denatured in a sample collection tube. Human papilloma virus hybrid capture hybrid capture assay gynecologic screening specimen transport medium these keywords were added by machine and not by the authors.
1170 797 985 40 429 492 1436 1163 626 1222 316 321 918 296 519 1353 521 1079 1113 383 1474 1239 326 953 30 597 1309 20 1160 862 591 449 810 770 1116 674 25 970 40 299 745 930 627 244 1298