It is provided a method of designing at least one oligonucleotide for nucleic acid detection comprising the following steps in any order: (I) identifying and/or selecting region(s) of at least one target nucleic acid to be amplified, the region(s) having an efficiency of amplification (AE) higher than the average AE; and (II) designing at least one oligonucleotide capable of hybridizing to the selected region(s). It is also provided a method of detecting at least one target nucleic acid comprising the steps of: (i) providing at least one biological sample; (ii) amplifying nucleic acid(s) comprised in the biological sample; (iii) providing at least one oligonucleotide capable of hybridizing to at least one target nucleic acid, if present in the biological sample; and (iv) contacting the oligonucleotide(s) with the amplified nucleic acids and detecting the oligonucleotide(s) hybridized to the target nucleic acid(s). In particular, the method is for detecting the presence of at least one pathogen, for example a virus, in at least one human biological sample. The probes may be placed on a support, for example a microarray.