Rapid Visual Detection of Hepatitis C Virus Using Reverse Transcription Recombinase-Aided Amplification–Lateral Flow Dipstick

Wang, Haili; Zhang, Yuhang; Zhou, Jingming; Li, Ming; Chen, Yumei; Liu, Yankai; Liu, Hongliang; Ding, Peiyang; Liang, Chao; Zhu, Xifang; Zhang, Ying; Cheng, Xin; Zhang, Gaiping; Wang, Aiping

doi:10.3389/fcimb.2022.816238

Cited by 16 publications

(9 citation statements)

References 35 publications

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“…MIRA-LFD assays have been employed in the detection of some human pathogens, such as SARS-CoV-2 and hepatitis C virus ( 35 – 37 ), but they have been rarely employed in the veterinary field. To our knowledge, this is the first study to report the detection of BCoV using this method, and this study shall inspire the development of MIRA-LFD assays for the detection of other human and animal viruses.…”

Section: Discussionmentioning

confidence: 99%

Development of a multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for bovine coronavirus detection

Feng²,

Sun³

et al. 2023

Front. Vet. Sci.

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Bovine coronavirus (BCoV) is a major cause of infectious disease in cattle, causing huge economic losses to the beef and dairy industries worldwide. BCoV can infect humans and multiple other species of animals. A rapid, reliable, and simple test is needed to detect BCoV infection in suspected farms. In this study, we developed a novel multienzyme isothermal rapid amplification (MIRA) and lateral flow dipstick (LFD) combination assay, targeting a highly conserved region of the viral nucleocapsid (N) gene for BCoV detection. The MIRA-LFD assay was highly specific and sensitive, comparable to a published reverse transcription quantitative PCR (RT-qPCR) assay for BCoV detection. Compared with the published RT-qPCR assay, the κ value of the MIRA-LFD assay in the detection of 192 cattle clinical samples was 0.982. The MIRA-LFD assay did not require sophisticated instruments and the results could be observed with eyes. Our results showed that the MIRA-LFD assay was a useful diagnostic tool for rapid on-site detection of BCoV.

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Section: Discussionmentioning

confidence: 99%

Development of a multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for bovine coronavirus detection

Feng²,

Sun³

et al. 2023

Front. Vet. Sci.

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“…Moreover, immunosuppressed infected individuals can have false-negative outcomes. In recent decades, reverse transcription polymerase chain reaction technology was used to identify HCV infection, which has good sensitivity and can detect viremia in acute infection during the window period (when HCV-Ab assay is still negative) [ 25 ]. However, this assay is inappropriate in low- and middle-income countries owing to the high costs of thermo-cycling instruments and need for trained technicians.…”

Section: Discussionmentioning

confidence: 99%

Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications

Shi,

Zhou,

Dong

et al. 2024

BMC Microbiol

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Objectives In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV. Methods A set of LAMP primer was designed according to 5’untranslated region (5’UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections. Results An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5’UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference. Conclusions These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions.

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“…While techniques such as immunoassay and qPCR play an important role in the diagnosis of HCV, rapid and accurate POC testing is important to identify individuals with HCV, particularly in resource-limited settings where access or availability of molecular testing is still limited. Therefore, to fill this gap, there is a need to develop a new molecular assay for the rapid detection of HCV RNA in resource-limited settings [ 97 ]. Recently, Kham-Kjing et al developed and evaluated a fast and sensitive method for the detection of HCV RNA.…”

Section: Potential Of Crispr/cas System In Viral Hepatitis Diagnosismentioning

confidence: 99%

Potential of CRISPR/Cas system as emerging tools in the detection of viral hepatitis infection

Bahrulolum

Tarrahimofrad

Rouzbahani

et al. 2023

Virol J

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Viral hepatitis, the most common cause of inflammatory liver disease, affects hundreds of millions of people worldwide. It is most commonly associated with one of the five nominal hepatitis viruses (hepatitis A–E viruses). HBV and HCV can cause acute infections and lifelong, persistent chronic infections, while HAV and HEV cause self-limiting acute infections. HAV and HEV are predominantly transmitted through the fecal-oral route, while diseases transmitted by the other forms are blood-borne diseases. Despite the success in the treatment of viral hepatitis and the development of HAV and HBV vaccines, there is still no accurate diagnosis at the genetic level for these diseases. Timely diagnosis of viral hepatitis is a prerequisite for efficient therapeutic intervention. Due to the specificity and sensitivity of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated sequences (Cas) technology, it has the potential to meet critical needs in the field of diagnosis of viral diseases and can be used in versatile point-of-care (POC) diagnostic applications to detect viruses with both DNA and RNA genomes. In this review, we discuss recent advances in CRISPR–Cas diagnostics tools and assess their potential and prospects in rapid and effective strategies for the diagnosis and control of viral hepatitis infection.

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Rapid Visual Detection of Hepatitis C Virus Using Reverse Transcription Recombinase-Aided Amplification–Lateral Flow Dipstick

Cited by 16 publications

References 35 publications

Development of a multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for bovine coronavirus detection

Development of a multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for bovine coronavirus detection

Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications

Potential of CRISPR/Cas system as emerging tools in the detection of viral hepatitis infection

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