A qPCR targeted against the viral replication origin designed to quantify total amount of filamentous phages and phagemids

dc.contributorUniversidad Nacional de Asunción - Facultad de Ciencias Químicas
dc.creatorMéndez Scolari, José Emilio
dc.creatorFlorentín Pavía, Marcos Marcelo
dc.creatorMujica, M. P.
dc.creatorRojas, N.
dc.creatorSotelo, Pablo Hernán
dc.date2022-04-27T23:38:00Z
dc.date2022-04-27T23:38:00Z
dc.date2019-03
dc.date.accessioned2023-09-25T13:30:08Z
dc.date.available2023-09-25T13:30:08Z
dc.identifierhttp://hdl.handle.net/20.500.14066/3732
dc.identifierhttps://doi.org/10.1007/s12088-019-00798-x
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8806550
dc.descriptionFilamentous bacteriophages are widely used in phage display technology. The most common quantification method is lysis plaque formation test (PFT). This technique has several disadvantages, and only quantifies infective phages and is not effective when phagemids are used. We developed a qPCR method directed against the M13 replication origin, which detects between 3.3 9 103 and 3.3 9 108 viral genome copies with a linearity of R2 = 0.9998. Using this method we were able to observe a difference of approximately ten more phages than with the PFT. This difference was not due to the presence of a free genome, which suggests the presence of non-infective particles. Using a DNaseI treatment, we observed the presence of 30% to 40% of unpackaged genome in recombinant phage modified in PIII or PVIII. The qPCR method with a DNase I treatment is an efficient method to quantify the total amount of filamentous phages.
dc.descriptionCONACYT – Consejo Nacional de Ciencia y Tecnología
dc.descriptionPROCIENCIA
dc.languageeng
dc.relationPINV15-224
dc.rightsopen access
dc.rights© Association of Microbiologists of India 2019
dc.subject6 Producción y tecnología industrial
dc.subjectPHAGE QUANTITATION
dc.subjectPHAGE DISPLAY
dc.subjectQPCR
dc.subjectPHAGEMIDS
dc.subjectFARMACOLOGIA
dc.titleA qPCR targeted against the viral replication origin designed to quantify total amount of filamentous phages and phagemids
dc.typeresearch article


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