Establishment of immunoassay for detecting HPV16 E6 and E7 RNA.
Cervical carcinoma is the most prevalent malignancy second only to breast cancer among women worldwide. Since more than 99% of cervical cancers are caused by human papilloma virus (HPV), measurement of HPV (HPV test) was commonly used in screening risk and/or early stage of cervical cancer as well as assessing the efficacies of the treatments that can decrease the incidence of cervical cancer. Many approaches that diagnose HPV infections have been developed, while most of them have distinct shortcomings. We here established a novel immunoassay method in which the pairs of unlabeled DNA probes firstly bind to HPV16 E6 and E7 RNAs to form the DNA-RNA hybrids, and the hybrids will subsequently be identified by S9.6 antibody. The sensitivity of this highly specific method can reach ~0.923 pg/mL and ~0.424 pg/mL of in vitro transcribed HPV16 E6 and E7 RNA, respectively, and reverse transcription and polymerase chain reaction (PCR) amplification were no longer needed. Thus, our immunoassay approaches can precisely reflect the actually viral load that is related to the course of HPV infection. In addition, it has also fast and low cost characteristic feature.
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210023, China.|MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, 210061, China;
Sen
S
Ding
Department of Pharmaceutical Sciences, North Dakota State University, Fargo, ND 58108-6050, United States.
Steven Y
SY
Qian
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210023, China.|MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, 210061, China;
Yang
Y
Zhang
Institute and Hospital of Stomatology, Nanjing University Medical School, Nanjing, 210093, China.
Wenlei
W
Wu
Nanjing Maternity and Child Health Care Hospital, Nanjing, 210004, China.
Gensheng
G
Lu
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210023, China.|MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, 210061, China;
Yan
Y
Lu
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210023, China.|MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, 210061, China;
Xiujing
X
Feng
School of Chemistry &Life Sciences, Nanjing University Jinling College, Nanjing, 210089, China.
Li
L
Li
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, 210023, China.|MOE Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing Biomedical Research Institute, Nanjing University, Nanjing, 210061, China;
Pingping
P
Shen
D004268:DNA-Binding Proteins / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D027383:Papillomaviridae / Q000276:immunology* / Q000302:isolation & purification*; D012367:RNA, Viral / Q000276:immunology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004268:DNA-Binding Proteins; C052603:E6 protein, Human papillomavirus type 18; D009856:Oncogene Proteins, Viral; D012367:RNA, Viral