Experimental data for Figs 2–7. Article Swipe
Jia-Rui Liang (21021470)
,
Shu-Ning Yan (21021473)
,
Han-Yin Yang (8484912)
,
Shuo Yang (209399)
,
Yu-Juan Shen (394322)
,
Le-Le Huo (21021476)
,
Yu-Chun Cai (115603)
,
Zi-Ran Mo (21021479)
,
Bin Zheng (45359)
,
Bin Xu (10691)
,
Wei Hu (6560)
·
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pntd.0013007.s005
YOU?
·
· 2025
· Open Access
·
· DOI: https://doi.org/10.1371/journal.pntd.0013007.s005
Background Necator americanus is the predominant species causing hookworm infections in humans. Despite advancements in prevention strategies, mild cases of infection still occur, highlighting the need for improved detection technology. Recombinase Polymerase Amplification (RPA) is an isothermal molecular diagnostic known for its sensitivity, speed, portability, and widespread application in detecting various pathogens. Although several molecular assays are available for N. americanus, they have limitations in detecting mild N. americanus infections. Methods Fluorescent RPA primers and probes targeting the N. americanus internal transcribed spacer 2 (ITS2) gene were developed. The method’s detection limit was assessed via serial dilution of genomic DNA. Specificity was confirmed against Clonorchis sinensis, Schistosoma japonicum, Fasciola hepatica, Ascaris lumbricoides, Enterobius vermicularis and Ancylostoma duodenale. Thirty samples identified as positive by Kato-Katz, along with 11 samples identified as negative by the method, were tested to evaluate the sensitivity and specificity of fluorescent RPA. Additionally, 287 field samples were tested for validation with these methods. All positive samples were identified as either N. americanus or A. duodenale. Results This study successfully developed a fluorescent RPA assay targeting the ITS2 gene of N. americanus. The length of the amplified fragment was 237 bp. Optimized conditions were achieved, resulting in a minimum detection limit of 1fg/µL, with no cross-reactivity with other pathogens. In laboratory validation, the fluorescent RPA assay demonstrated 100% sensitivity (30/30) and 100% specificity (11/11) compared to the Kato-Katz, and 100% sensitivity (29/29) and 91.7% specificity (11/12) when compared to the semi-nested PCR. In field validation using human fecal samples, the fluorescent RPA assay showed a sensitivity of 90.0% (36/40) and a specificity of 91.1% (225/247) compared to the Kato-Katz. And the sensitivity of the fluorescent RPA method compared to the semi-nested PCR method was 100% (34/34), while the specificity was 90.5% (229/252). Conclusions The fluorescent RPA assay presents a rapid and dependable method for detecting N. americanus in fecal samples. Its high sensitivity and specificity provide significant utility for field surveillance and early identification of N. americanus infections. This advancement could facilitate the rapid molecular diagnosis of N. americanus disease in hookworm-endemic regions.
Related Topics
Concepts
Serial dilution
Molecular biology
Biology
Detection limit
Assay sensitivity
Enterobius
Recombinase Polymerase Amplification
Loop-mediated isothermal amplification
Fluorescence
Polymerase chain reaction
Real-time polymerase chain reaction
Gene
Virology
Amplicon
Ascaris
DNA extraction
Chromatography
Feces
Metadata
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- OA Status
- green
- OpenAlex ID
- https://openalex.org/W7111342535
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W7111342535Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1371/journal.pntd.0013007.s005Digital Object Identifier
- Title
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Experimental data for Figs 2–7.Work title
- Type
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datasetOpenAlex work type
- Publication year
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2025Year of publication
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2025-04-08Full publication date if available
- Authors
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Jia-Rui Liang (21021470), Shu-Ning Yan (21021473), Han-Yin Yang (8484912), Shuo Yang (209399), Yu-Juan Shen (394322), Le-Le Huo (21021476), Yu-Chun Cai (115603), Zi-Ran Mo (21021479), Bin Zheng (45359), Bin Xu (10691), Wei Hu (6560)List of authors in order
- Open access
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YesWhether a free full text is available
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greenOpen access status per OpenAlex
- Concepts
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Serial dilution, Molecular biology, Biology, Detection limit, Assay sensitivity, Enterobius, Recombinase Polymerase Amplification, Loop-mediated isothermal amplification, Fluorescence, Polymerase chain reaction, Real-time polymerase chain reaction, Gene, Virology, Amplicon, Ascaris, DNA extraction, Chromatography, FecesTop concepts (fields/topics) attached by OpenAlex
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0Total citation count in OpenAlex
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| abstract_inverted_index.positive | 122, 158 |
| abstract_inverted_index.presents | 300 |
| abstract_inverted_index.regions. | 345 |
| abstract_inverted_index.samples, | 251 |
| abstract_inverted_index.samples. | 312 |
| abstract_inverted_index.(225/247) | 267 |
| abstract_inverted_index.Optimized | 194 |
| abstract_inverted_index.achieved, | 197 |
| abstract_inverted_index.amplified | 189 |
| abstract_inverted_index.available | 58 |
| abstract_inverted_index.confirmed | 103 |
| abstract_inverted_index.detecting | 50, 66, 307 |
| abstract_inverted_index.detection | 29, 91, 202 |
| abstract_inverted_index.developed | 173 |
| abstract_inverted_index.diagnosis | 338 |
| abstract_inverted_index.hepatica, | 110 |
| abstract_inverted_index.infection | 21 |
| abstract_inverted_index.molecular | 38, 55, 337 |
| abstract_inverted_index.resulting | 198 |
| abstract_inverted_index.sinensis, | 106 |
| abstract_inverted_index.targeting | 77, 178 |
| abstract_inverted_index.(229/252). | 294 |
| abstract_inverted_index.<i>Necator | 2 |
| abstract_inverted_index.Background | 1 |
| abstract_inverted_index.Enterobius | 113 |
| abstract_inverted_index.Kato-Katz, | 124, 230 |
| abstract_inverted_index.Kato-Katz. | 271 |
| abstract_inverted_index.Polymerase | 32 |
| abstract_inverted_index.conditions | 195 |
| abstract_inverted_index.dependable | 304 |
| abstract_inverted_index.developed. | 88 |
| abstract_inverted_index.diagnostic | 39 |
| abstract_inverted_index.facilitate | 334 |
| abstract_inverted_index.identified | 120, 129, 161 |
| abstract_inverted_index.infections | 10 |
| abstract_inverted_index.isothermal | 37 |
| abstract_inverted_index.japonicum, | 108 |
| abstract_inverted_index.laboratory | 213 |
| abstract_inverted_index.method’s | 90 |
| abstract_inverted_index.pathogens. | 52, 211 |
| abstract_inverted_index.prevention | 16 |
| abstract_inverted_index.validation | 153, 247 |
| abstract_inverted_index.widespread | 47 |
| abstract_inverted_index.<i>ITS2</i> | 180 |
| abstract_inverted_index.Conclusions | 295 |
| abstract_inverted_index.Fluorescent | 72 |
| abstract_inverted_index.Recombinase | 31 |
| abstract_inverted_index.Schistosoma | 107 |
| abstract_inverted_index.Specificity | 101 |
| abstract_inverted_index.advancement | 332 |
| abstract_inverted_index.application | 48 |
| abstract_inverted_index.fluorescent | 144, 175, 216, 253, 277, 297 |
| abstract_inverted_index.infections. | 70, 330 |
| abstract_inverted_index.limitations | 64 |
| abstract_inverted_index.predominant | 6 |
| abstract_inverted_index.semi-nested | 243, 283 |
| abstract_inverted_index.sensitivity | 140, 221, 233, 258, 274, 315 |
| abstract_inverted_index.significant | 319 |
| abstract_inverted_index.specificity | 142, 225, 237, 264, 291, 317 |
| abstract_inverted_index.strategies, | 17 |
| abstract_inverted_index.technology. | 30 |
| abstract_inverted_index.transcribed | 82 |
| abstract_inverted_index.validation, | 214 |
| abstract_inverted_index.advancements | 14 |
| abstract_inverted_index.demonstrated | 219 |
| abstract_inverted_index.highlighting | 24 |
| abstract_inverted_index.portability, | 45 |
| abstract_inverted_index.sensitivity, | 43 |
| abstract_inverted_index.successfully | 172 |
| abstract_inverted_index.surveillance | 323 |
| abstract_inverted_index.(<i>ITS2)</i> | 85 |
| abstract_inverted_index.<i>Clonorchis | 105 |
| abstract_inverted_index.Additionally, | 146 |
| abstract_inverted_index.Amplification | 33 |
| abstract_inverted_index.lumbricoides, | 112 |
| abstract_inverted_index.<i>Ancylostoma | 116 |
| abstract_inverted_index.americanus</i> | 3, 69, 80, 165, 309, 329, 341 |
| abstract_inverted_index.duodenale</i>. | 117, 168 |
| abstract_inverted_index.identification | 326 |
| abstract_inverted_index.americanus</i>, | 61 |
| abstract_inverted_index.americanus</i>. | 184 |
| abstract_inverted_index.cross-reactivity | 208 |
| abstract_inverted_index.hookworm-endemic | 344 |
| abstract_inverted_index.vermicularis</i> | 114 |
| cited_by_percentile_year | |
| countries_distinct_count | 0 |
| institutions_distinct_count | 11 |
| citation_normalized_percentile |