Development of an improved PCR–ICT hybrid assay for direct detection of Legionellae and Legionella pneumophila from cooling tower water specimens

Horng, Yu-Tze; Soo, Po-Chi; Shen, Bin-Jon; Hung, Y.-Y.; Lo, Kai-Yin; Su, Hsun-Pi; Wei, Jun-Rong; Hsieh, Shang-Chen; Hsueh, Po-Ren; Lai, Hsin‐Chih

doi:10.1016/j.watres.2006.03.033

Cited by 24 publications

(17 citation statements)

References 35 publications

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“…Commercial and inhouse-developed NAATs are used in both clinical and environmental laboratories; however, the list of Legionella genes amplified is limited. The most common targets include a conserved segment of the rRNA genes for the 5S and 16S subunits, the 16S-23S spacer, and/or the macrophage inhibitor protein mip, found primarily in the genus Legionella and highly conserved in all L. pneumophila isolates (226,(332)(333)(334)(349)(350)(351)(352)(353)(354)(355)(356)(357)(358). Several studies have also examined alternative chromosomal targets, such as dotA, gyrB, dnaJ, wzm, and wzt (340).…”

Section: Nucleic Acid-based Molecular Diagnosticsmentioning

confidence: 99%

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

2015

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SUMMARY Legionnaires' disease (LD) is an often severe and potentially fatal form of bacterial pneumonia caused by an extensive list of Legionella species. These ubiquitous freshwater and soil inhabitants cause human respiratory disease when amplified in man-made water or cooling systems and their aerosols expose a susceptible population. Treatment of sporadic cases and rapid control of LD outbreaks benefit from swift diagnosis in concert with discriminatory bacterial typing for immediate epidemiological responses. Traditional culture and serology were instrumental in describing disease incidence early in its history; currently, diagnosis of LD relies almost solely on the urinary antigen test, which captures only the dominant species and serogroup, Legionella pneumophila serogroup 1 (Lp1). This has created a diagnostic “blind spot” for LD caused by non-Lp1 strains. This review focuses on historic, current, and emerging technologies that hold promise for increasing LD diagnostic efficiency and detection rates as part of a coherent testing regimen. The importance of cooperation between epidemiologists and laboratorians for a rapid outbreak response is also illustrated in field investigations conducted by the CDC with state and local authorities. Finally, challenges facing health care professionals, building managers, and the public health community in combating LD are highlighted, and potential solutions are discussed.

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Section: Nucleic Acid-based Molecular Diagnosticsmentioning

confidence: 99%

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

2015

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“…The increased incidence of nosocomial Legionella outbreaks in recent years has led to the development of efficient molecular methods for the identification of free and amoeba-associated Legionella spp. (Miyamoto et al, 1997;Horng et al, 2006). In addition to legionellosis, other diseases produced by amoebaresistant pathogens, such as those caused by Mycobacterium spp.…”

Section: Introductionmentioning

confidence: 99%

A new pentaplex-nested PCR to detect five pathogenic bacteria in free living amoebae

Calvo¹,

Gregorio²,

Garcı́a³

et al. 2013

Water Research

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“…The MBLF technology has ever been also applied on the detection of various Polymerase Chain Reaction (PCR) products, for instances, allergens [13], [14], bacteria [15], pathogens [16], antibodies [17], and antigens [18], [19]. Our recent study revealed that an electric field could enhance the detection performance of single-trended Deoxyribonucleic acids (DNA) on the MBLF strips [20].…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of Gene HLA-A3101 on Membrane Based Lateral-Flow Strips

Wu¹,

Huang²

2018

IJBBB

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Gene Human leukocyte antigen (HLA)-A3101 is known responsible of a medicine allergy which may cause a fatal outcome. General clinics need a quick test to determine if a patient is engaged with the related gene defect and avoid the medical prescription from suspicious drugs. In this study, Gene HLA-A3101 is detected on membrane-based lateral-flow (MBFL) strips, incorporated with an immune reaction and color readouts developed by gold nanoparticles. The HLA-A-3101 DNA modified with biotin and digoxigenin labels was amplified by the Polymerase Chain Reaction (PCR) and then detected on the MBFL strips. The optimal condition of antibody concentration was at least 10µg/ml, to avoid the antibody-gold complex from aggregation. The signal can be observed as low as 0.49 ng of the PCR product by naked eyes. The specificity of PCR primers was also investigated their annealing validity. The detection specificity from other species genes, including a plant, a virus, and a human gene, were also verified. The coefficient of variation values of the intra assay, which tests the same sample in the same day, and the inter assay, which tests three samples in three consecutive days, are in satisfactory ranges of 4.6% and 6.5%, respectively.

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Development of an improved PCR–ICT hybrid assay for direct detection of Legionellae and Legionella pneumophila from cooling tower water specimens

Cited by 24 publications

References 35 publications

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

Current and Emerging Legionella Diagnostics for Laboratory and Outbreak Investigations

A new pentaplex-nested PCR to detect five pathogenic bacteria in free living amoebae

Rapid Detection of Gene HLA-A3101 on Membrane Based Lateral-Flow Strips

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