Usefulness of capillary electrophoresis-based multiplex PCR assay for species-specific identification of Candida spp.

Mallus, Francesca; Martis, Silvia; Serra, C; Loi, G.; Camboni, Tania

doi:10.1016/j.mimet.2012.11.022

Cited by 6 publications

(6 citation statements)

References 11 publications

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“…and Mallus et al . [ 34 , 35 ]. Both studies used the Seegene Seeplex PCR assay (Seegene Diagnostic, Seoul, South Korea) for nucleotide amplification, but CGE was carried out on two different platforms QIAxcel (Qiagen, Hilden, Germany) [ 35 ] and the MultiNA (Shimadzu Corp., Tokyo, Japan) [ 34 ], respectively.…”

Section: Discussionmentioning

confidence: 99%

“…[ 34 , 35 ]. Both studies used the Seegene Seeplex PCR assay (Seegene Diagnostic, Seoul, South Korea) for nucleotide amplification, but CGE was carried out on two different platforms QIAxcel (Qiagen, Hilden, Germany) [ 35 ] and the MultiNA (Shimadzu Corp., Tokyo, Japan) [ 34 ], respectively. Our assay here differs as SCGE relies on fluorescent-labeled primers for PCR amplification whereas QIAxcel and MultiNA devices are adapted for analysis of conventional PCR products amplified by non-labeled primers requiring an additional step to identify nucleic acid with e.g.…”

Section: Discussionmentioning

confidence: 99%

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Sequencer-Based Capillary Gel Electrophoresis (SCGE) Targeting the rDNA Internal Transcribed Spacer (ITS) Regions for Accurate Identification of Clinically Important Yeast Species

et al. 2016

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Accurate species identification of Candida, Cryptococcus, Trichosporon and other yeast pathogens is important for clinical management. In the present study, we developed and evaluated a yeast species identification scheme by determining the rDNA internal transcribed spacer (ITS) region length types (LTs) using a sequencer-based capillary gel electrophoresis (SCGE) approach. A total of 156 yeast isolates encompassing 32 species were first used to establish a reference SCGE ITS LT database. Evaluation of the ITS LT database was then performed on (i) a separate set of (n = 97) clinical isolates by SCGE, and (ii) 41 isolates of 41 additional yeast species from GenBank by in silico analysis. Of 156 isolates used to build the reference database, 41 ITS LTs were identified, which correctly identified 29 of the 32 (90.6%) species, with the exception of Trichosporon asahii, Trichosporon japonicum and Trichosporon asteroides. In addition, eight of the 32 species revealed different electropherograms and were subtyped into 2–3 different ITS LTs each. Of the 97 test isolates used to evaluate the ITS LT scheme, 96 (99.0%) were correctly identified to species level, with the remaining isolate having a novel ITS LT. Of the additional 41 isolates for in silico analysis, none was misidentified by the ITS LT database except for Trichosporon mucoides whose ITS LT profile was identical to that of Trichosporon dermatis. In conclusion, yeast identification by the present SCGE ITS LT assay is a fast, reproducible and accurate alternative for the identification of clinically important yeasts with the exception of Trichosporon species.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Sequencer-Based Capillary Gel Electrophoresis (SCGE) Targeting the rDNA Internal Transcribed Spacer (ITS) Regions for Accurate Identification of Clinically Important Yeast Species

et al. 2016

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“…The Seeplex assay was found to be specific and rapid for identifying six clinically important Candida species [75] and 80 clinical strains of Candida spp. [76]. Gago et al [77] developed PCR-high resolution melting (HRM) analysis to genotype Candida albicans strains and compared the resulting data with those from microsatellite length polymorphism (MLP)-CE analysis.…”

Section: Detecting and Diagnosing Fungimentioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

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“…Multiplex PCR has been used to detect Candida spp. (Mallus et al 2013;Vahidnia et al 2015) and Aspergillus spp. (Amini et al 2015;Logotheti et al 2009) as has real-time PCR (Emam & Abd El-salam 2015;Horváth et al 2013).…”

Section: Introductionmentioning

confidence: 99%

“…(Amini et al 2015;Logotheti et al 2009) as has real-time PCR (Emam & Abd El-salam 2015;Horváth et al 2013). Several PCR techniques have targeted ribosomal DNA of Candida (Cerikçioğlu et al 2010;Mallus et al 2013;Than et al 2012) and Aspergillus (Walsh et al 2011). Although these PCR methods have been useful for the identification of fungal species, they either only identify species within a particular genus or detect the fungus at genus level.…”

Section: Introductionmentioning

confidence: 99%

Dual Panel Multiplex PCR Assay for Rapid Detection of Medically Important Fungi and Resistant Species of Candida and Aspergillus

Jainlabdin¹,

Chua²,

Nizam³

et al. 2018

JSM

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Usefulness of capillary electrophoresis-based multiplex PCR assay for species-specific identification of Candida spp.

Cited by 6 publications

References 11 publications

Sequencer-Based Capillary Gel Electrophoresis (SCGE) Targeting the rDNA Internal Transcribed Spacer (ITS) Regions for Accurate Identification of Clinically Important Yeast Species

Sequencer-Based Capillary Gel Electrophoresis (SCGE) Targeting the rDNA Internal Transcribed Spacer (ITS) Regions for Accurate Identification of Clinically Important Yeast Species

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Dual Panel Multiplex PCR Assay for Rapid Detection of Medically Important Fungi and Resistant Species of Candida and Aspergillus

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