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Recombinase polymerase amplification - lateral flow dipstick for rapid and visual detection of Blastocystis spp.
Xuefang Mei1, Changwei Su1, Jiahui Xin1,2
1Xinxiang Key Laboratory of Pathogenic Biology, Department of Pathogenic Biology, School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, Henan, China.
Abstract:
Blastocystis spp. is a ubiquitous protozoon in the intestinal tract of human and many animals. Microscopic examination is the main method of clinical diagnosis for Blastocystis spp., which is prone to false negative. A simple and rapid diagnosis of Blastocystis spp. infection is an important step to prevent and control blastocystosis. Here, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay was developed for rapid visual detection of Blastocystis spp. DNA amplification could be performed within 18 min at 37°C. The minimum DNA detection limit was 1 pg/μL, and there was no cross-reactivity with 12 other non-target pathogens, which was consistent with the sensitivity of conventional PCR (cPCR). Furthermore, 56 fecal samples from the Third Affiliated Hospital of Xinxiang Medical University were tested using RPA and cPCR methods respectively, and the results were completely consistent. The results show that RPA-LFD method has high accuracy and visual results, which provides a new choice for the differential diagnosis and rapid field detection of Blastocystis spp.
Insights
A new recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay offers rapid and accurate visual detection of Blastocystis spp. DNA. This method provides a reliable alternative for diagnosing Blastocystis infections, overcoming limitations of traditional microscopic examination.
Area of Science:
- Medical Parasitology
- Molecular Diagnostics
- Microbiology
Background:
- Blastocystis spp. is a common intestinal protozoon.
- Current diagnostic methods like microscopy are prone to false negatives.
- Rapid and accurate diagnosis is crucial for controlling blastocystosis.
Purpose of the Study:
- To develop a rapid, visual, and accurate diagnostic assay for Blastocystis spp.
- To evaluate the performance of the developed assay compared to conventional methods.
Main Methods:
- Development of a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay.
- Testing the assay's DNA detection limit and specificity against non-target pathogens.
- Validation using 56 fecal samples compared against conventional PCR (cPCR).
Main Results:
- The RPA-LFD assay achieved DNA amplification within 18 minutes at 37°C.
- The minimum DNA detection limit was 1 pg/μL with no cross-reactivity.
- Results from 56 clinical samples showed complete consistency with cPCR.
Conclusions:
- The developed RPA-LFD assay is highly accurate and provides visual results.
- This assay offers a valuable new option for rapid field detection and differential diagnosis of Blastocystis spp. infections.
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