title: Alternative molecular tests for virological diagnosis.
abstract: Several nucleic acid amplification techniques (NAATs), particularly PCR and real-time PCR, are currently used in the routine clinical laboratories. Such approaches have allowed rapid diagnosis with a high degree of sensitivity and specificity. However, conventional PCR methods have several intrinsic disadvantages such as the requirement for temperature cycling apparatus, and sophisticated and costly analytical equipments. Therefore, amplification at a constant temperature is an attractive alternative method to avoid these requirements. A new generation of isothermal amplification techniques are gaining a wide popularity as diagnostic tools due to their simple operation, rapid reaction and easy detection. The main isothermal methods reviewed here include loop-mediated isothermal amplification, nucleic acid sequence-based amplification, and helicase-dependent amplification. In this review, design criteria, potential of amplification, and application of these alternative molecular tests will be discussed and compared to conventional NAATs.
authors.affiliation: Department of Public Health and Microbiology, University Hospital San Giovanni Battista di Torino, University of Turin, Turin, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Sidoti
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Bergallo
authors.forename: Cristina
authors.initials: C
authors.lastname: Costa
authors.forename: Rossana
authors.initials: R
authors.lastname: Cavallo
mesh_terms: D004265:DNA Helicases / Q000032:analysis / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012327:RNA Virus Infections / Q000175:diagnosis*; D012328:RNA Viruses / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D004265:DNA Helicases