Detection of HCV RNA by the asymmetric gap ligase chain reaction.

Marshall, R L; Laffler, Thomas G.; Cerney, M B; Sustachek, J C; Kratochvil, Jon; Morgan, Roy

doi:10.1101/gr.4.2.80

Cited by 26 publications

(14 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Both LCR and isothermal amplification were first described in 1989 [89,90]. Although these techniques are well known and used for viral load detection in infection with, for example, HIV and hepatitis C virus, they are not used for the detection or identification of bacteria in blood cultures [91][92][93][94]. LCR has only been used for the detection of Mycobacterium tuberculosis in respiratory specimens, and the isothermal transcription-mediated technique has only been used for the rapid identification of Candida spp.…”

Section: Multiplex Assaysmentioning

confidence: 99%

Conventional and molecular techniques for the early diagnosis of bacteraemia

Paolucci

Landini

Sambri

2010

International Journal of Antimicrobial Agents

View full text Add to dashboard Cite

Bloodstream infections account for 30-40% of all cases of severe sepsis and septic shock, and are major causes of morbidity and mortality. Diagnosis of bloodstream infections must be performed promptly so that adequate antimicrobial therapy can be started and patient outcome improved. An ideal diagnostic technology would identify the infecting organism(s) and their determinants of antibiotic resistance, in a timely manner, so that appropriate pathogen-driven therapy could begin promptly. Unfortunately, despite the essential information it provides, blood culture, the gold standard, largely fails in this purpose because time is lost waiting for bacterial or fungal growth. Several efforts have been made to optimize the performance of blood culture, such as the development of technologies to obtain rapid detection of microorganism(s) directly in blood samples or in a positive blood culture. The ideal molecular method would analyze a patient's blood sample and provide all the information needed to immediately direct optimal antimicrobial therapy for bacterial or fungal infections. Furthermore, it would provide data to assess the effectiveness of the therapy by measuring the clearance of microbial nucleic acids from the blood over time. None of the currently available molecular methods is sufficiently rapid, accurate or informative to achieve this. This review examines the principal advantages and limitations of some traditional and molecular methods commercially available to help the microbiologist and the clinician in the management of bloodstream infections.

show abstract

Section: Multiplex Assaysmentioning

confidence: 99%

Conventional and molecular techniques for the early diagnosis of bacteraemia

Paolucci

Landini

Sambri

2010

International Journal of Antimicrobial Agents

View full text Add to dashboard Cite

show abstract

“…In addition, NAT exhibits a very high sensitivity of detection, serves as an independent method for confirming infection in indeterminate serology cases, helps to discriminate between chronic and resolved HCV cases, and confirms infections in HIV-and HCV-seropositive newborns. Several technologies for the detection of HIV and/or HCV nucleic acids are currently available, such as branched DNA signal amplification (10,12,18,39), nucleic acid sequence-based amplification (40,42), strand displacement amplification (29,44), the ligase chain reaction (24,27), transcription-mediated amplification (21,28), and PCR (1,25,31,38). The main shortcomings of the first-generation NAT technologies, rendering them unsuitable for high-throughput screening, include technical difficulty, significant hands-on time, lengthy incubation times, large sample volume requirements, high cost, relatively high rate of false positives, and inconsistency in detection of genetic variants (11,17,33,34).…”

mentioning

confidence: 99%

Highly Sensitive Multiplex Assay for Detection of Human Immunodeficiency Virus Type 1 and Hepatitis C Virus RNA

Giachetti¹,

Linnen²,

Kolk³

et al. 2002

J Clin Microbiol

105

122

View full text Add to dashboard Cite

Various nucleic acid assays have been developed and implemented for diagnostics and therapeutic monitoring of human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) infections. The highthroughput, semiautomated assays described here were developed to provide a method suitable for screening plasma specimens for the presence of HIV-1 and HCV RNAs. Three assays were developed: a multiplex HIV-1/HCV assay for simultaneous detection of HIV-1 and HCV, and discriminatory assays for specific detection of HIV-1 and HCV. The assay systems utilize three proprietary technologies: (i) target capture-based sample preparation, (ii) transcription-mediated amplification (TMA), and (iii) hybridization protection assay (HPA). An internal control is incorporated into each reaction to control for every step of the assay and identify random false-negative reactions. The assays demonstrated a sensitivity of at least 100 copies/ml for each target, and they detected with similar sensitivity all major variants of HCV and HIV-1, including HIV-1 group O strains. Assay sensitivity for one virus was not affected by the presence of the other. The specificity of these TMA-driven assays was >99.5% in both normal donor specimens and plasma containing potentially interfering substances or other blood-borne pathogens. Statistical receiver operating characteristic plots of 1 ؊ specificity versus sensitivity data determined very wide analyte cutoff values for each assay at the point at which the assay specificity and sensitivity were both >99.5%. The sensitivity, specificity, and throughput capability predict that these assays will be valuable for large-volume plasma screening, either in a blood bank setting or in other diagnostic applications.

show abstract

“…Most studies on gLCR have used oligonucleotides with similar T m's although oligonucleotides with considerably different Tm's have also been used successfully (Grimberg et ai., 1993;Marshall et al, 1994), and could result in higher amplification efficiency (P. Lin, unpublished data). The "extending" oligonucleotide X or Z (Fig.…”

Section: The Gap Ligase Chain Reactionmentioning

confidence: 99%

“…One of the reaction design challenges lies in the fact that the reverse transcription (RT) reaction that generates cDNA from an RNA template typically uses four dNTPs for polymerization while gLCR uses three or less dNTPs for gap-filling. Marshall et al (1994) have reported a modification of gLCR called asymmetric gap LCR (AGLCR) for the detection of hepatitis C virus (HCV) RNA that uses three dNTPs and four oligonucleotides. The gap region in (2 and 11 nucleotides, respectively).…”

Section: Amplification Of Specific Target Rna Sequences Using Glcrmentioning

confidence: 99%

Technologies for Detection of DNA Damage and Mutations

Pfeifer¹

1996

View full text Add to dashboard Cite

Detection of HCV RNA by the asymmetric gap ligase chain reaction.

Cited by 26 publications

References 17 publications

Conventional and molecular techniques for the early diagnosis of bacteraemia

Conventional and molecular techniques for the early diagnosis of bacteraemia

Highly Sensitive Multiplex Assay for Detection of Human Immunodeficiency Virus Type 1 and Hepatitis C Virus RNA

Technologies for Detection of DNA Damage and Mutations

Contact Info

Product

Resources

About