Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples

Jiang, Xian; Dong, Derong; Bian, Lihong; Zou, Dayang; He, Xiaoming; Ao, Da; Yang, Zhan; Huang, Simo; Liu, Ningwei; Liu, Wei; Huang, Liuyu

doi:10.3389/fmicb.2016.00916

Cited by 37 publications

(22 citation statements)

References 29 publications

(31 reference statements)

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“…All eight PSR primer sets either failed to generate amplicons or displayed non-specific amplification in water controls when designed following the instructions from the original publication. However, follow-up publications described enhancements to PSR by adding additional inner primers (S1 Fig) [39,40]. Implementation of this modification enabled successful amplification for one set of PSR primers (S2C Fig).…”

Section: Primer Design and Selectionmentioning

confidence: 99%

Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics

2020

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A number of isothermal DNA amplification technologies claim to be ideal for point-of-need (PON) applications as they enable reactions to be performed using a single-temperature heat source (e.g. water bath). Thus, we examined several isothermal amplification methods focusing on simplicity, cost, sensitivity and reproducibility to identify the most suitable method(s) for low resource PON applications. A number of methods were found unsuitable as they either involved multiple temperature incubations, were relatively expensive or required relatively large amounts target DNA for amplification. Among the methods examined, loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) were found to be the most suitable for PON applications as they are both single step methods that provide highly sensitive and reproducible amplifications. The speed of LAMP reactions was greatly enhanced, up to 76%, with the addition of loop primers while the presence of swarm primers and the sequestration of free magnesium ions with nucleotides also enhanced the amplification speed. In contrast, we were unable to enhance RPA's performance from the original published literature. While both RPA and LAMP have some drawbacks, either isothermal technology can reliably be used for on-site diagnostics with minimal equipment.

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Section: Primer Design and Selectionmentioning

confidence: 99%

Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics

2020

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“…The developed assay shows some similarities with LAMP, (CPA) 15 17 , nucleic acid sequence dependent amplification (NASBA) 18 , rolling circle replication (RCR) 19 and recently polymerase spiral reaction (PSR) described for the first time by Liu et al . 20 then subsequently applied by other scientists for detection of Candida albicans 21 . All these methods have an advantage to overcome the requirement of cycling which is compulsory in case of PCR-based techniques or real-time PCR.…”

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confidence: 99%

Polymerase cross-linking spiral reaction (PCLSR) for detection of African swine fever virus (ASFV) in pigs and wild boars

Woźniakowski

Frączyk

Kowalczyk

et al. 2017

Sci Rep

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The study reports the development of a polymerase cross-linking spiral reaction (PCLSR) for the detection of African swine fever virus (ASFV) DNA in blood collected from infected pigs and wild boars. The method uses 3 specifically designed primers. Two outer-spiral primers comprising of 3′ sequences complementary to ASFV p72 gene sequence and 5′end sequences complementary to exogenous gene of black widow alpha-latrotoxin as well as additional ASFV specific cross-linking primer. The method is specific exclusively to ASFV DNA without cross-reactions with cDNA of classical swine fever virus (CSFV), porcine reproductive respiratory syndrome (PRRSV) or porcine epidemic diarrhea virus (PEDV). The sensitivity of this technique reached 7.2 × 102 copies per μl−1 of plasmid containing p72 gene. The PCLSR was conducted at 65 °C creating cross-linked complex structures. The results of PCLSR were visualized using SYBR Green I dye, gel electrophoresis while the reaction progress was traced using real-time PCR system that resulted in registration of fluorescent curves and melting peaks at 85.3 °C. The developed PCLSR was examined using blood or tissue samples collected from selected 17 ASF cases from infected wild boars and 3 outbreaks in pigs. Further tests have been also conducted using 55 tissue samples from 23 outbreaks and 22 cases. These results showed that PCLSR might be further used for preliminary and cost-effective detection and surveillance of ASFV.

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“…We invented PSR 10 , 11 , a new in vitro isothermal nucleic acid amplification method, and own all the intellectual property rights. In this method, only two primers were required, each with the same forward sequence at the 5′ end, or one with a forward sequence and the other with a reverse sequence at the 5′ end, enabling the primers to use themselves as templates to substantially replicate the target nucleic acid segment.…”

Section: Preparation Of the M Tuberculosis Dementioning

confidence: 99%

Development and application of a rapid Mycobacterium tuberculosis detection technique using polymerase spiral reaction

Liu¹,

Zou²,

He³

et al. 2018

Sci Rep

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Mycobacterium tuberculosis is an age-old bacterium that is difficult to eliminate. A simple and rapid diagnostic method is of great importance to prevent the spread of M. tuberculosis. Here, we developed a low-cost rapid M. tuberculosis nucleic acid detection technique, named GenePop, which enabled the storage and transport of M. tuberculosis diagnostic reagent at ambient temperatures, without the need for professional operations or expensive instrumentation. Using a vitrification method, we vitrified heat-unstable components onto the cap of a reaction tube, and placed heat-stable components at the bottom of the reaction tube by sealing them with paraffin wax. The all-in-one detection tube, when used together with our other invention—a multi-functional sample treatment tube pre-filled with a nucleic acid-releasing agent—only required three simple steps to yield results. A comparative analysis with a commercial qPCR kit for M. tuberculosis indicated that our new technique had a concordance rate of 91.6%, showing no cross-reactivity with 11 other bacteria. The complete operation time was only 65 min. It is suitable for use in field settings or by personnel in grass-root units, and is applicable in household activities, hence can be used in developing countries.

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Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples

Cited by 37 publications

References 29 publications

Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics

Evaluation and improvement of isothermal amplification methods for point-of-need plant disease diagnostics

Polymerase cross-linking spiral reaction (PCLSR) for detection of African swine fever virus (ASFV) in pigs and wild boars

Development and application of a rapid Mycobacterium tuberculosis detection technique using polymerase spiral reaction

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