Cervical carcinoma is the second most common cancer in women worldwide with greater than 99% of the cases caused by human papillomaviruses (HPVs). Early detection of HPVs especially the high risk types (HR-HPVs) are essential to prevent the disease progression. The existing methods for HPV detection, such as qPCR are of high sensitivity and specificity, but the need for expensive machinery and well-trained personnel slow down the disease detection. The emerging Cas12a-based method presents a new technique for nucleic acid detection. However, it is time-consuming and labor-intensive when used for HPV detection, as several reactions are required in order to identify multiple HPV infections. We herein present a non-genotyping method for 13 types of HR-HPV detection in a single reaction by combining the isothermal recombinase polymerase amplification (RPA) method with CRISPR-Cas12a technology. The result could be achieved in 35 min with high sensitivity (500 copies per reaction). This assay represents great advances for the application of RPA-Cas12a system and holds a great potential to address the key challenges facing the HPV diagnostics.

Background: To investigate the results and analyze the diagnostic value of the combined application of EBUS-TBNA and T-SPOT in patients with granuloma in mediastinal-hilar lymph nodes. Methods: Patients who underwent EBUS-TBNA and pathologically confirmed as lymph node granulomatous lesions from January 2020 to May 2021 were retrospectively recruited. The baseline clinical characteristics, tuberculosis T cell spot test, pathological and imaging results of these subjects were collected. The common disease accompanied by granulomas lesion in mediastinal-hilar lymph nodes were analyzed and compared. Results: Our results represented that among 60 patients with lymph node granulomatosis, caseous necrosis was only detected in 2 cases, 13 cases with positive acid-resistant staining were diagnosed as lymphoid tuberculosis, however, none of them manifested the typical symptoms of tuberculosis poisoning. 5 cases were diagnosed as sarcoidosis, and oral prednisone acetate tablets were effective. T-SPOT test results showed a positive result in 26 cases, we noticed that the positive acid-resistant staining cases (T-SPOT detection in 9 cases) had higher antigen values of T-SPOT than the other 17 cases with positive T-SPOT (P＜0.01) Conclusion:EBUS-TBNA combined T-SPOT test has great significance to differentiate lymphoid tuberculosis and sarcoidosis, helping promote the diagnostic rate of granulomas lesion in mediastinal-hilar lymph nodes.

Cervical carcinoma is the second most common cancer in women worldwide with greater than 99% of the cases caused by human papillomaviruses (HPVs). Early detection of HPVs especially the high risk types (HR-HPVs) are essential to prevent the disease progression. The existing methods for HPV detection, such as qPCR are of high sensitivity and specificity, but the need for expensive machinery and well-trained personnel slow down the disease detection. The emerging Cas12a-based method presents a new technique for nucleic acid detection. However, it is time-consuming and labor-intensive when used for HPV detection, as several reactions are required in order to identify multiple HPV infections. We herein present a non-genotyping method for 13 types of HR-HPV detection in a single reaction by combining the isothermal recombinase polymerase amplification (RPA) method with CRISPR-Cas12a technology. The result could be achieved in 30 minutes with high sensitivity (500 copies per reaction). This assay reprsents great advances for the application of RPA-Cas12a system and holds a great potential to address the key challenges facing the HPV diagnostics.