A portable multiplex biosensor was developed for detection and genotyping of parvovirus B19 DNA, exploiting lensless CL imaging. The reaction chip is composed of a polydimethylsiloxane microfluidic layer coupled with a glass slide on which oligonucleotide probes specific for three different B19 genotypes are covalently immobilized in a 3?×?3 array. The reaction chip was used in hybridization reactions with biotin-labeled targets and then hybrids were then detected by means of an avidin-HRP conjugate, upon addition of a CL substrate for HRP