Development and application of multiprobe real-time PCR method targeting the hsp65 gene for differentiation of Mycobacterium species from isolates and sputum specimens.

Journal of Clinical Microbiology
Kijeong KimBum-Joon Kim

Abstract

We developed a multiprobe real-time PCR assay targeting hsp65 (HMPRT-PCR) to detect and identify mycobacterial isolates and isolates directly from sputum specimens. Primers and probes for HMPRT-PCR were designed on the basis of the hsp65 gene sequence, enabling the recognition of seven pathogenic mycobacteria, including Mycobacterium tuberculosis, M. avium, M. intracellulare, M. kansasii, M. abscessus, M. massiliense, and M. fortuitum. This technique was applied to 24 reference and 133 clinical isolates and differentiated between all strains with 100% sensitivity and specificity. Furthermore, this method was applied to sputum specimens from 117 consecutive smear-positive patients with smear results of from a trace to 3+. These results were then compared to those obtained using the rpoB PCR-restriction analysis method with samples from cultures of the same sputum specimens. The HMPRT-PCR method correctly identified the mycobacteria in 89 samples (76.0%, 89/117), and moreover, the sensitivity level was increased to 94.3% (50/53) for sputa with an acid-fast bacillus score equal to or greater than 2+. Our data suggest that this novel HMPRT-PCR method could be a promising approach for detecting pathogenic mycobacterial species from ...Continue Reading

References

Aug 13, 1992·Nature·B R Bloom
Aug 21, 1992·Science·B R Bloom, C J Murray
Feb 1, 1992·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·G MassenkeilR Lüthy
Jul 1, 1992·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·E Wolinsky
Jun 6, 1991·The New England Journal of Medicine·P F BarnesD E Snider
Sep 1, 1993·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·D HuminerS D Pitlik
Dec 10, 2003·Journal of Clinical Microbiology·Toïdi AdékambiMichel Drancourt
Dec 31, 2004·Infection·D Wagner, L S Young
Jul 15, 2005·International Journal of Systematic and Evolutionary Microbiology·Hong KimBum-Joon Kim
Dec 20, 2005·Journal of Korean Medical Science·Won-Jung KohKyung Soo Lee
Jan 19, 2006·Clinical Microbiology Reviews·M J EspyT F Smith
Jul 26, 2008·Diagnostic Microbiology and Infectious Disease·Bum-Joon KimJae-Joon Yim
Dec 20, 2008·The Journal of Molecular Diagnostics : JMD·Suporn FoongladdaRuengpung Sutthent

❮ Previous
Next ❯

Citations

Feb 11, 2012·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Fumihiko KitagawaKoji Otsuka
Dec 24, 2011·Future Microbiology·Juan Carlos Palomino
Sep 14, 2012·Mediterranean Journal of Hematology and Infectious Diseases·Fathiah ZakhamRajae Elaouad
Nov 26, 2013·Human Immunology·Seema D ShekhawatRajpal S Kashyap
Nov 22, 2013·BioMed Research International·Sara MacenteRosilene Fressatti Cardoso
Feb 27, 2015·Journal of Applied Microbiology·J-U KimH K An
Mar 15, 2014·Revista Do Instituto De Medicina Tropical De São Paulo·Yvana Maria Maia de AlbuquerqueVera Magalhães
Jul 15, 2016·Journal of Clinical Microbiology·Gianny P ScoleriGeraint B Rogers
Feb 14, 2012·Optometry and Vision Science : Official Publication of the American Academy of Optometry·Xiaowei LiuQin Long
Nov 17, 2012·Expert Review of Molecular Diagnostics·Max Maurin
Jan 22, 2011·Molecular Diagnosis & Therapy·Sathish SankarGopalan Sridharan
Apr 25, 2015·British Journal of Biomedical Science·J A O'ConnorB Lucey
Jun 22, 2019·The European Respiratory Journal·Steven CowmanMichael R Loebinger
Nov 12, 2020·Revista Da Sociedade Brasileira De Medicina Tropical·Aline Dos Santos PeixotoZulma Maria Medeiros

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.