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Circoviruses belong to the family Circoviridae, which is classified into two genera, Circovirus and Cycloviru s. Some circoviruses have been identified in various organisms and/or their fecal samples and might be associated with diseases in their hosts. However, few circoviruses are reported in human. Recently, two novel circoviruses HCirV-1 and HCirV-2 were identified in humans through next-generation sequencing and were defined as the species of Circovirus human in the family Circoviridae . Both viruses are suspected to be associated with liver diseases, particularly among immunosuppressed people, showing potential health implication. Investigation on the prevalence of both viruses, and identification of vulnerable population are important and need a rapid, accurate, and specific assay for detecting and screening the two viruses. In this study, we developed a duplex HiFi-LAMP assay for simultaneous detection of HCirV-1 and HCirV-2. The assay exhibits high sensitivity with LOD of 64 and 49 copies per 25 µL reaction for HCirV-1 and HCirV-2, respectively. The duplex assay was demonstrated to have a rapid reaction time within 35 min. Clinical screening tests showed that neither HCirV-1 nor HCirV-2 were detected among 875 patients with infection of HBV, HIV-1 or other viruses. Large-scale screening of both viruses in diverse populations is encouraged to enhance our understanding of their relevance to various diseases. Supplementary Information The online version contains supplementary material available at 10.1186/s12879-024-10283-6.
Circoviruses belong to the family Circoviridae, which is classified into two genera, Circovirus and Cycloviru s. Some circoviruses have been identified in various organisms and/or their fecal samples and might be associated with diseases in their hosts. However, few circoviruses are reported in human. Recently, two novel circoviruses HCirV-1 and HCirV-2 were identified in humans through next-generation sequencing and were defined as the species of Circovirus human in the family Circoviridae . Both viruses are suspected to be associated with liver diseases, particularly among immunosuppressed people, showing potential health implication. Investigation on the prevalence of both viruses, and identification of vulnerable population are important and need a rapid, accurate, and specific assay for detecting and screening the two viruses. In this study, we developed a duplex HiFi-LAMP assay for simultaneous detection of HCirV-1 and HCirV-2. The assay exhibits high sensitivity with LOD of 64 and 49 copies per 25 µL reaction for HCirV-1 and HCirV-2, respectively. The duplex assay was demonstrated to have a rapid reaction time within 35 min. Clinical screening tests showed that neither HCirV-1 nor HCirV-2 were detected among 875 patients with infection of HBV, HIV-1 or other viruses. Large-scale screening of both viruses in diverse populations is encouraged to enhance our understanding of their relevance to various diseases. Supplementary Information The online version contains supplementary material available at 10.1186/s12879-024-10283-6.
The impact of porcine circovirus (PCV) on the worldwide pig industry is profound, leading to notable economic losses. Early and prompt identification of PCV is essential in managing and controlling this disease effectively. A range of detection techniques for PCV have been developed and primarily divided into two categories focusing on nucleic acid or serum antibody identification. The methodologies encompass conventional polymerase chain reaction (PCR), real-time fluorescence quantitative PCR (qPCR), fluorescence in situ hybridization (FISH), loop-mediated isothermal amplification (LAMP), immunofluorescence assay (IFA), immunohistochemistry (IHC), and enzyme-linked immunosorbent assay (ELISA). Despite their efficacy, these techniques are often impeded by the necessity for substantial investment in equipment, specialized knowledge, and intricate procedural steps, which complicate their application in real-time field detections. To surmount these challenges, a sensitive, rapid, and specific PCV detection method using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas12a/13a coupled with isothermal amplification, such as enzymatic recombinase amplification (ERA), recombinase polymerase amplification (RPA), and loop-mediated isothermal amplification (LAMP), has been developed. This novel method has undergone meticulous optimization for detecting PCV types 2, 3, and 4, boasting a remarkable sensitivity to identify a single copy per microliter. The specificity of this technique is exemplary, with no observable interaction with other porcine viruses such as PEDV, PRRSV, PRV, and CSFV. Its reliability has been validated with clinical samples, where it produced a perfect alignment with qPCR findings, showcasing a 100% coincidence rate. The elegance of merging CRISPR-Cas technology with isothermal amplification assays lies in its on-site testing without the need for expensive tools or trained personnel, rendering it exceptionally suitable for on-site applications, especially in resource-constrained swine farming environments. This review assesses and compares the process and characteristics inherent in the utilization of ERA/LAMP/RPA-CRISPR-Cas12a/Cas13a methodologies for the detection of PCV, providing critical insights into their practicality and effectiveness.
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