High sensitivity, loop-mediated isothermal amplification combined with colorimetric gold-nanoparticle probes for visual detection of high risk human papillomavirus genotypes 16 and 18

Kumvongpin, Ratchanida; Jearanaikool, Patcharee; Wilailuckana, Chotechana; Sae-ung, Nattaya; Prasongdee, Prinya; Daduang, Sakda; Wongsena, Metee; Boonsiri, Patcharee; Kiatpathomchai, Wansika; Swangvaree, Sukumarn; Sandee, Alisa; Daduang, Jureerut

doi:10.1016/j.jviromet.2016.04.008

Cited by 59 publications

(39 citation statements)

References 17 publications

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“…LFD is sufficient for the detection of hybridized LAMP products (21,22), and the results are easy to read without the use of carcinogens, such as ethidium bromide. LAMP detection methods have been successfully developed for HPV16 and HPV18 using visual turbidity and gel electrophoresis, and these assays can be completed within 70 and 115 min, respectively (15). In the present study, the same LAMP primer sets were used as previously published, but LAMP-LFD combined detection provided higher sensitivity and was completed within a shorter time period compared with a previous report (8).…”

Section: Hpv16 Hpv18 ------------------------------------------------mentioning

confidence: 79%

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Detection assay for HPV16 and HPV18 by loop-mediated isothermal amplification with lateral flow dipstick tests

Kumvongpin

Jearanaikoon

Wilailuckana

et al. 2017

Molecular Medicine Reports

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Cervical cancer is the third highest cause of death in developing countries and most commonly results from high‑risk human papillomavirus (HR‑HPV) infection. Among HR‑HPV genotypes, HPV16 and HPV18 are the most prevalent in cervical cancers. Therefore, the present study aimed to develop a detection assay for HPV16 and HPV18 infection using loop‑mediated isothermal amplification (LAMP) with lateral flow dipstick (LFD) tests. This assay is a simplified, user‑friendly method for the visual detection of HPV genotypes. DNA was extracted from clinical tissue samples, and HPV genotyping was performed using nested polymerase chain reaction (PCR). The clinical samples were demonstrated to include 44 HPV16‑positive, 18 HPV18‑positive and 80 HPV‑negative samples. All DNA samples were also used as templates for a LAMP reaction (30 min at 65˚C), and subsequently, a fluorescein isothiocyanate‑labelled probe was hybridized with the reaction product. Finally, the LFD test was performed. The sensitivity of the LAMP‑LFD test was higher than LAMP‑turbidity, exhibiting up to 100‑fold higher sensitivity for HPV16 and 10‑fold higher sensitivity for HPV18. All HPV16 and HPV18‑positive samples generated positive results in both assays; however, 22 samples detected as HPV‑negative by LAMP‑turbidity exhibited positive results by LAMP‑LFD test (22 of 80 samples). Therefore, these samples were further examined using quantitative (q)PCR. The results demonstrated that 20 out of the 22 samples designated positive by LAMP‑LFD, but negative by LAMP turbidity, gave a positive result with qPCR, while the remaining 2 samples were negative by qPCR. The present results suggested that LAMP‑LFD provided higher sensitivity than LAMP‑turbidity and nested PCR. Thus, the LAMP‑LFD test developed in the present study might be useful for the detection of HPV16 and HPV18 in local hospitals.

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Section: Hpv16 Hpv18 ------------------------------------------------mentioning

confidence: 79%

“…Evaluation of clinical samples. All 142 clinical samples were examined using LAMP-turbidity (15) and LAMP-LFD assays. The sensitivity and specificity of these assays were calculated using standard formulae based on the results of nested PCR.…”

Section: Sensitivity Of Lamp-turbidity and Lamp-lfd Assaysmentioning

confidence: 99%

Detection assay for HPV16 and HPV18 by loop-mediated isothermal amplification with lateral flow dipstick tests

Kumvongpin

Jearanaikoon

Wilailuckana

et al. 2017

Molecular Medicine Reports

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“…As been discussed in this paper, LAMP could be one of the diagnostic method alternative in relation to its rapidity, sensitivity (Ishiguro et al 2015;Kumvongpin et al 2016;Besuschio et al 2017) and specificity. The most significant advantage of LAMP would be its rapidity where LAMP allows immediate diagnosis.…”

Section: Advantages Of Lampmentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

et al. 2018

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“…We chose instead to develop a method to extract specific target sequences from such tissue, so as to increase the reliability, accuracy, and sensitivity of PCR analysis. While HPV DNA can be specifically detected and amplified for diagnostic purposes using Oligo-AuNP probes [26,27], to our knowledge no protocols have been previously reported for the type-specific extraction of HPV DNA from FFPE tissue.…”

Section: Introductionmentioning

confidence: 99%

A high yield gold nanoparticle-based DNA isolation method for human papillomaviruses genotypes from cervical cancer tissue samples

Seyyedi¹,

Farjadian²,

Farhadi³

et al. 2019

Preprint

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Gold nanoparticles (AuNPs) are commonly used in biosensors of various kinds. The purification of DNA from cancer tissues is an important step in diagnostic and therapeutic development, but current methods are not optimal. Many cervical cancer patients are also susceptible to high-risk human papillomavirus (HR-HPV) infection. Accurate viral diagnosis has so far relied on the extraction of adequate amounts of DNA from formalin-fixed, paraffin-embedded (FFPE) tissue samples. Since the sensitivity and specificity of commercially available purification kits are not optimal, we designed a DNA purification method based on AuNPs to purify sufficient amounts of HR-HPV DNA from cervical cancer tissue samples. AuNPs were coated with a series of oligonucleotide probes to hybridize to specific DNA sequences of HR-HPV genotypes. With this method, we recovered 733 out of 800 copies of type-specific HPV DNA with complete specificity, compared to 36 copies with a standard commercial kit (Qiagen FFPE).

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High sensitivity, loop-mediated isothermal amplification combined with colorimetric gold-nanoparticle probes for visual detection of high risk human papillomavirus genotypes 16 and 18

Cited by 59 publications

References 17 publications

Detection assay for HPV16 and HPV18 by loop-mediated isothermal amplification with lateral flow dipstick tests

Detection assay for HPV16 and HPV18 by loop-mediated isothermal amplification with lateral flow dipstick tests

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

A high yield gold nanoparticle-based DNA isolation method for human papillomaviruses genotypes from cervical cancer tissue samples

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