Evidence map›Paper›PMID 39893430›Full record

ArticleVirology journal2025

Development of a duplex real-time recombinase aided amplification assay for the simultaneous and rapid detection of PCV3 and PCV4.

Renjie Sun, Hanze Liu, Siqi Sun, Yating Wang, Ying Shan, Xiaoliang Li, Weihuan Fang, Yongle Yang, Ronghui Xie, Lingyan Zhao

Abstract readEvaluation Study
In one paragraph

Article in Virology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Rapid and simultaneous detection ofFrontiers in cellular and infection microbiology · 2026
    Article
  2. Review
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Renjie SunZhejiang Provincial Center for Animal Disease Control and Prevention, Hangzhou, 311199, China.
Hanze LiuChina Animal Health and Epidemiology Center, Qingdao, 266032, China.
Siqi SunZhejiang Provincial Center for Animal Disease Control and Prevention, Hangzhou, 311199, China.
Yating WangZhejiang Provincial Center for Animal Disease Control and Prevention, Hangzhou, 311199, China.
Ying ShanInstitute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
Xiaoliang LiInstitute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
Weihuan FangInstitute of Preventive Veterinary Medicine and Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, Zhejiang University, Hangzhou, 310058, China.
Yongle YangXianghu Laboratory, Hangzhou, 311231, China. yang-yl@zju.edu.cn.
Ronghui XieZhejiang Provincial Center for Animal Disease Control and Prevention, Hangzhou, 311199, China. hunter429@tom.com.
Lingyan ZhaoZhejiang Provincial Center for Animal Disease Control and Prevention, Hangzhou, 311199, China. zhaoly96@163.com.

Funding

National Natural Science Foundation of China 32172819The High-level Talents Special Support Plan of Zhejiang Province 2021R52041Zhejiang Province Science and Technology Cooperation Project of "Three Rural and Nine Parties" 2023SNJF059Zhejiang Provincial Natural Science Foundation of China LQ23C18001
6 · The paper itself

Abstract

backgroundPorcine circoviruses 3 (PCV3) and 4 (PCV4) are emerging pathogens with global implications for swine industry, disturbing the diagnosis of PCVs associated diseases due to a range of similar clinical symptoms and increasingly coinfections. A rapid and accurate method for detection of PCV3 and PCV4 is critical for controlling the transmission of associated disease.

methodsWe developed a duplex real-time recombinase aided amplification (RAA) assay for detection of both PCV3 and PCV4 simultaneously. The assay was completed within 20 min at 39℃ with the designed optimal primers and probes.

resultsThe established assay was more convenient and simpler operation compared with conventional molecular biological assays. The assay achieved a detection limit of 73.67 copies/reaction for each circovirus (at 95% probability by probit regression analysis) and showed high specificity and no cross-reactivity with other important porcine viruses (including PCV2). The intra- and inter-group coefficients of variation (CV) were ranged from 2.08 to 4.97%, indicating high stability and reliability. Comparative analysis with PCV3 and PCV4 qPCR on 60 clinical samples and artificially spiked samples indicated high congruence (the kappa value was 0.966 and 1, respectively, with p < 0.001), with only minor discrepancies, validating effectiveness of the duplex RAA assay in detecting co-infections and its suitability for preliminary clinical diagnosis of PCV3 and PCV4.

conclusionsThis study provides a robust basis for multiplex detection of veterinary pathogens using RAA technique, enhancing the field's capacity to control PCV3 and PCV4, and supporting reliable aid for epidemiological understanding of emerging circoviruses.

Indexed as

Circoviridae InfectionsCircovirusMolecular Diagnostic TechniquesNucleic Acid Amplification TechniquesReal-Time Polymerase Chain ReactionSwine DiseasesAnimalsDNA PrimersRecombinasesReproducibility of ResultsSensitivity and SpecificitySwineDNA PrimersRecombinasesDuplex detectionPorcine circoviruses 3 (PCV3)Porcine circoviruses 4 (PCV4)Recombinase aided amplification (RAA)

Identifiers

PMID39893430
PMCPMC11786481

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.