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

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 100 and 101 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.

The effect of reduced RDI of chemotherapy on the outcome of breast cancer patients.

The aims of this study were to investigate the impact of the relative dose intensity (RDI) of chemotherapy on disease-free survival (DFS) and overall survival (OS), to identify the optimal RDI cut-off points with the docetaxel, epirubicin and cyclophosphamide (TEC) regimen for stage I-III breast cancer patients and to explore the adverse events in these patients. To achieve this, we performed a retrospective analysis of breast cancer patients treated at the First Affiliated Hospital of Chongqing Medical University in 2011. The results showed that among 293 patients with the TEC regimen, 85% and 80% were the cut-off points at which a high RDI was associated with better overall survival (HR = 2.04; 95% CI 1.13, 3.70; p = 0.02) and disease-free survival (HR = 1.97; 95% CI 1.14-3.42; p = 0.02), respectively. Among 169 HR(+) patients, 80% was the cut-off point for DFS (HR = 2.33; 95% CI 1.07-5.08; p = 0.03), and 85% was the cut-off point for OS (HR = 3.00; 95% CI 1.24-7.26; p = 0.02). Among 105 HR(-) patients, 80% was the cut-off point for OS (HR = 2.86; 95% CI 1.05-7.80; p = 0.04). Of 293 patients, neutropenia, nausea, and vomiting were found to be correlated with the level of RDI. In conclusion, a higher RDI of chemotherapy is associated with better survival but with a higher probability of causing adverse events. To optimize survival benefits, the RDI should be maintained ≥ 85% for HR(+) patients and ≥ 80% for HR(-) patients.