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This systematic review aimed to examine biological AOD testing methods from hospital\u2010based studies of injured patients and quantify what proportion reported key information on those testing methods.<\/jats:p><\/jats:sec><jats:sec><jats:title>Approach<\/jats:title><jats:p>Observational studies published in English from 2010 onwards involving biological AOD testing for injured patients presenting to hospital were included. Studies examining single injury causes were excluded. Extracted data included concentration thresholds for AOD detection (e.g., lower limits of detection, author\u2010defined cut\u2010offs), test type (e.g., immunoassay, breathalyser) and approach (e.g., routine, clinical discretion), timing of testing, sample type and the proportion of injured cases tested for AODs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Findings<\/jats:title><jats:p>Of 83 included studies, 76 measured alcohol and 37 other drugs. Forty\u2010nine studies defined blood alcohol concentration thresholds (ranging from 0 to 0.1\u2009g\/100\u2009mL). Seven studies defined concentration thresholds for other drugs. Testing approach was reported in 39\/76 alcohol and 18\/37 other drug studies. Sample type was commonly reported (alcohol: <jats:italic>n<\/jats:italic>\u2009=\u200969\/76; other drugs: <jats:italic>n<\/jats:italic>\u2009=\u200928\/37); alcohol was typically measured using blood (<jats:italic>n<\/jats:italic>\u2009=\u200960) and other drugs using urine (<jats:italic>n<\/jats:italic>\u2009=\u200920). Studies that reported the proportion of cases tested (alcohol: <jats:italic>n<\/jats:italic>\u2009=\u200953\/76; other drugs: <jats:italic>n<\/jats:italic>\u2009=\u200928\/37), reported that between 0% and 89% of cases were not tested for alcohol and 0% and 91% for other drugs. Timing of testing was often unreported (alcohol: <jats:italic>n<\/jats:italic>\u2009=\u200961; other drugs: <jats:italic>n<\/jats:italic>\u2009=\u200930).<\/jats:p><\/jats:sec><jats:sec><jats:title>Implications and Conclusion<\/jats:title><jats:p>Variation in AOD testing methods alongside incomplete reporting of those methods limits data comparability and interpretation. 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