title: Four DNA extraction methods used in loop-mediated isothermal amplification for rapid adenovirus detection.
abstract: Loop-mediated isothermal amplification (LAMP) assays have become powerful tools for rapid diagnosis of infectious diseases. A more efficient, convenient and cheaper method for template preparation from the pellets or supernatants of nasopharyngeal aspirates was sought. Three DNA extraction methods (boiling, boiling in 1% Triton X-100, and treating with 0.02M NaOH) were compared with the commonly used DNAzol DNA extraction method. DNA preparations were then subjected to adenovirus (ADV) detection using LAMP assays and 119 clinical samples. The specificities for all three methods were 100% compared with the DNAzol method. The sensitivity of the boiling method was greater than that for the other two methods. The templates extracted from supernatants of nasopharyngeal aspirates using the boiling technique were further evaluated. Higher sensitivity (90.9%) and specificity (96.5%) were observed for LAMP assays compared with those from quantitative PCR assays. In conclusion, for template preparation, boiling supernatants of nasopharyngeal aspirates had comparable sensitivity and specificity with the DNAzol method. There were the added advantages that the boiling technique was simpler, cheaper, and had a shorter processing time. The boiling technique could become a suitable substitute for the DNAzol method when LAMP assays are used for ADV detection.
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China. Electronic address: linqingz525@163.com.
authors.forename: Linqing
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Runan
authors.initials: R
authors.lastname: Zhu
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Deng
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Li
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Yaxin
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Run
authors.initials: R
authors.lastname: Tian
authors.affiliation: Laboratory of Virology, Capital Institute of Pediatrics, Beijing 100020, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qian
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification*; D000258:Adenovirus Infections, Human / Q000175:diagnosis* / Q000821:virology; D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009305:Nasopharynx / Q000821:virology; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: Adenoviruses; DNA extraction; Loop-mediated isothermal amplification (LAMP); Pediatric patients