A comparative study of three different nucleic acid amplification techniques combined with microchip electrophoresis for HPV16 E6/E7 mRNA detection.
Research towards nucleic acid amplification technologies for detection of human papillomavirus (HPV) 16 E6/E7 mRNA was carried out in combination with microchip electrophoresis (MCE). The approaches of nucleic acid sequence based amplification (NASBA), one-step RT-PCR and two-step RT-PCR were successfully developed. NASBA was a simple enzymatic reaction, which directly amplified HPV16 mRNA by isothermal amplification, leaving out the complex and tedious operation. One-step RT-PCR simplified the amplification step, while two-step RT-PCR was more sensitive and less vulnerable to the interference. Furthermore, instead of gel electrophoresis, microchip electrophoresis (MCE) for RNA assay was employed to realize high-throughput and rapid analysis. Finally, the results show that PCR-based or NASBA-based mRNA tests are valuable for HPV mRNA assay, which can be potentially applied for clinical diagnosis and prognosis of cervical and other anogenital carcinoma.
State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing 100029, China.
Quanli
Q
Liu
Xuexia
X
Lin
Luyao
L
Lin
Linglu
L
Yi
Haifang
H
Li
Jin-Ming
JM
Lin
D002021:Buffers; D046209:Electrophoresis, Microchip / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D050725:Papillomavirus E7 Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification; D012097:Repressor Proteins / Q000235:genetics*
D002021:Buffers; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16