Withatanung P.Vanaporn M.Chantratita N.Chareonsudjai S.Muangsombut V.Janesomboon S.Asawalertsaeng T.Clokie M.R.J.Galyov E.E.Korbsrisate S.Mahidol University2026-02-172026-02-172026-03-20Iscience Vol.29 No.3 (2026)https://repository.li.mahidol.ac.th/handle/123456789/115109Conventional diagnostic approaches involve identifying the bacterial pathogen from patient samples, which can delay results and miss low-bacteremia cases. Herein, we demonstrate that Burkholderia phages circulating in patient blood can serve as a powerful diagnostic biomarker. Building on this, a multiplex PCR targeting the Burkholderia pseudomallei phage terminase gene was developed to improve melioidosis diagnosis. The assay provided 97.2% sensitivity and 100% specificity, with results obtainable within 6 h. The strength of this PCR assay is the amplification of four different multicopy Burkholderia phage terminase genes from serum samples, resulting in increased sensitivity under low-bacteremia conditions. To our knowledge, this study reports a phage-based PCR assay that detects phage DNA directly from melioidosis patients’ blood, representing a shift from molecular pathogen-based to phage-based diagnostics. This approach not only improves sensitivity but also opens avenues for integrating phage biology into the diagnostic landscape of infectious diseases.MultidisciplinaryArticleSCOPUS10.1016/j.isci.2026.1148312-s2.0-10502969745925890042