Development of field-applicable endogenous internally controlled recombinase-aided amplification (EIC-RAA) assays for the detection of human papillomavirus genotypes 6 and 11 using sample releasing agent

He, Anna; Fang, Cheng; Ming, Yue; Tan, He; Zhang, Mingjie; Zhang, Ruiqing; Li, Jingyi; Nie, Min; Li, Fengyu; Hu, Yaxin; Shen, Xinxin; Rong, Xiuge

doi:10.1016/j.heliyon.2022.e11323

Cited by 2 publications

(1 citation statement)

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“…Moreover, in this study, we used the column extraction method to acquire the DNA templates of samples. Simpler methods for nucleic acid extraction, such as nucleic acid releasing agents, have been successfully applied and show the advantages of simplifying the operation process and saving time [ 42 , 43 ]. Our future work will focus on optimizing the procedure of nucleic acid extraction and rendering the entire process of detection more feasible for POCT.…”

Section: Discussionmentioning

confidence: 99%

Development of Enzymatic Recombinase Amplification Assays for the Rapid Visual Detection of HPV16/18

Ding,

Qi,

et al. 2023

J. Microbiol. Biotechnol.

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Human papillomavirus (HPV) types 16 and 18 are the major causes of cervical lesions and are associated with 71% of cervical cancer cases globally. However, public health infrastructures to support cervical cancer screening may be unavailable to women in low-resource areas. Therefore, sensitive, convenient, and cost-efficient diagnostic methods are required for the detection of HPV16/18. Here, we designed two novel methods, real-time ERA and ERA-LFD, based on enzymatic recombinase amplification (ERA) for quick point-of-care identification of the HPV E6/E7 genes. The entire detection process could be completed within 25 min at a constant low temperature (35–43°C), and the results of the combined methods could be present as the amplification curves or the bands presented on dipsticks and directly interpreted with the naked eye. The ERA assays evaluated using standard plasmids carrying the E6/E7 genes and clinical samples exhibited excellent specificity, as no cross-reaction with other common HPV types was observed. The detection limits of our ERA assays were 10 0 and 10 1 copies/μl for HPV16 and 18 respectively, which were comparable to those of the real-time PCR assay. Assessment of the clinical performance of the ERA assays using 114 cervical tissue samples demonstrated that they are highly consistent with real-time PCR, the gold standard for HPV detection. This study demonstrated that ERA-based assays possess excellent sensitivity, specificity, and repeatability for HPV16 and HPV18 detection with great potential to become robust diagnostic tools in local hospitals and field studies.

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Section: Discussionmentioning

confidence: 99%