A rapid and simple isothermal nucleic acid amplification test for detection of herpes simplex virus types 1 and 2

Kim, Hyun‐Jin; Tong, Yanhong; Tang, Wen; Quimson, Louisito; Cope, V.; Pan, Xiaojing; Motré, Aurélie; Kong, Richard Yuen Chong; Hong, Jian; Kohn, Debbie; Miller, Nancy S.; Poulter, Melinda D.; Kong, Huimin; Tang, Yi‐Wei; Yen‐Lieberman, Belinda

doi:10.1016/j.jcv.2010.09.006

Cited by 52 publications

(52 citation statements)

References 15 publications

(17 reference statements)

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“…The carboxyfluorescein-and biotin-labeled amplification-hybridization product migrates by capillary action through the next section of the LFS, where it binds the streptavidingold conjugate (detection probe) via the biotin label. As this complex reaches the detection band, it is captured there by anti-fluorescein isothiocyanate (FITC) antibodies (capture probe), resulting in a visible line (13). Noncaptured complexes and excess detection probe are stopped further downstream at the control line, providing a visual check of efficient flow along the stripe.…”

Section: Methodsmentioning

confidence: 99%

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

et al. 2012

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Reported methods for the detection of the yellow fever viral genome are beset by limitations in sensitivity, specificity, strain detection spectra, and suitability to laboratories with simple infrastructure in areas of endemicity. We describe the development of two different approaches affording sensitive and specific detection of the yellow fever genome: a real-time reverse transcriptionquantitative PCR (RT-qPCR) and an isothermal protocol employing the same primer-probe set but based on helicase-dependent amplification technology (RT-tHDA). Both assays were evaluated using yellow fever cell culture supernatants as well as spiked and clinical samples. We demonstrate reliable detection by both assays of different strains of yellow fever virus with improved sensitivity and specificity. The RT-qPCR assay is a powerful tool for reference or diagnostic laboratories with real-time PCR capability, while the isothermal RT-tHDA assay represents a useful alternative to earlier amplification techniques for the molecular diagnosis of yellow fever by field or point-of-care laboratories.

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Section: Methodsmentioning

confidence: 99%

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

et al. 2012

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“…tHDA technology (1) was previously applied to the detection of several infectious organisms (10,12,15). Specific modifications made to the technology for efficient multiplex amplification and detection were described by Doseeva et al (6a), which enable combined amplification and detection of three targets and an amplification control in a single reaction.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Chlamydia trachomatis and Neisseria gonorrhoeae Detection in Urine, Endocervical, and Vaginal Specimens by a Multiplexed Isothermal Thermophilic Helicase-Dependent Amplification (tHDA) Assay

et al. 2011

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We have developed a new research assay that combines sequence-specific sample preparation and isothermal amplification for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae infections. The assay targets both the omp gene and the cryptic plasmid of C. trachomatis and the multicopy opa gene of N. gonorrhoeae, which are amplified and detected in a single reaction. We evaluated the ability of the assay to detect C. trachomatis and N. gonorrhoeae infections in first-catch urine, swab, and liquid-based cytology samples. Total agreement between the new assay and APTIMA Combo 2 varied between 95.3% and 100%, depending on the sample type and target detected. Total agreement between the new assay and BD ProbeTec varied between 96.7% and 100%, depending on the sample type and target detected. The assay has a simple work flow, and endpoint results can be achieved in 3 h, including sample preparation. The assay described here was evaluated for research use and was compared to commercially available assays.Chlamydia trachomatis and Neisseria gonorrhoeae are among the most common causes of sexually transmitted infections (3). Infection with these organisms is often asymptomatic, which hinders diagnosis or treatment. Persistent infection can have serious negative consequences, including pelvic inflammatory disease and infertility. Untreated and undiagnosed infections also contribute to the further spread of the disease. Screening programs are recommended to prevent these consequences (17). Of the various testing methods currently available, nucleic acid amplification tests are among the most sensitive and specific methods for detection of C. trachomatis and N. gonorrhoeae organisms in clinical specimens (7, 9). The high sensitivity of nucleic acid amplification tests allows them to be used with less-invasive clinical samples, such as first-catch urine, which facilitates establishment of screening programs.The assay described here combines sequence-specific sample preparation and isothermal amplification for the detection of C. trachomatis and N. gonorrhoeae infections. Sequence-specific sample preparation is based on antibody capture of DNA-RNA hybrids. Ribo-oligonucleotide probes hybridize to specific target sequences, and the resulting hybrids are captured by antibodies specific for DNA-RNA hybrids. This process enriches the desired nucleic acid sequences in the sample. Antibodies are coupled to magnetic beads. Target amplification is accomplished by thermophilic helicase-dependent amplification (tHDA) (16). Amplification of captured strands of the DNA target occurs directly on magnetic beads through the combined action of primers, thermostable DNA polymerase, and helicase. Target DNA is detected by use of dually labeled probes in either endpoint or real-time detection modes. C. trachomatis and N. gonorrhoeae targets are amplified and detected simultaneously in a single closed reaction vessel.We investigated the ability of the assay to detect C. trachomatis and N. gonorrhoeae infections in clinical specimens. R...

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“…This maintains the ability to utilize these assays without sophisticated instrumentation but also allows the detection of multiple targets in a single reaction. A test developed to detect and differentiate herpes simplex virus 1 (HSV-1) and HSV-2 using this approach has demonstrated 100% sensitivity compared to viral culture, with a limit of detection as low as 5.5 copies per reaction (59). Further, this test could be performed on oral and genital cutaneous or mucocutaneous sources without the need for nucleic acid extraction and could be completed within 75 min.…”

Section: Singleplex Nucleic Acid Testsmentioning

confidence: 99%

Emerging Technologies for the Clinical Microbiology Laboratory

2014

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SUMMARY In this review we examine the literature related to emerging technologies that will help to reshape the clinical microbiology laboratory. These topics include nucleic acid amplification tests such as isothermal and point-of-care molecular diagnostics, multiplexed panels for syndromic diagnosis, digital PCR, next-generation sequencing, and automation of molecular tests. We also review matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) and electrospray ionization (ESI) mass spectrometry methods and their role in identification of microorganisms. Lastly, we review the shift to liquid-based microbiology and the integration of partial and full laboratory automation that are beginning to impact the clinical microbiology laboratory.

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A rapid and simple isothermal nucleic acid amplification test for detection of herpes simplex virus types 1 and 2

Cited by 52 publications

References 15 publications

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

Advanced Yellow Fever Virus Genome Detection in Point-of-Care Facilities and Reference Laboratories

Evaluation of Chlamydia trachomatis and Neisseria gonorrhoeae Detection in Urine, Endocervical, and Vaginal Specimens by a Multiplexed Isothermal Thermophilic Helicase-Dependent Amplification (tHDA) Assay

Emerging Technologies for the Clinical Microbiology Laboratory

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