Use of molecular methods to detect Shigella and infer phenotypic resistance in a Shigella treatment study.

Molecular diagnostic methods improve detection of Shigella yet their ability to detect Shigella drug resistance on direct stool specimens is less clear. We tested 673 stools from a Shigella treatment study in Bangladesh, including 154 culture-positive stools and their paired Shigella isolate. We utilized a TaqMan Array Card that included qPCR assays for 24 enteropathogens and 36 antimicrobial resistance (AMR) genes. Shigella was detected by culture in 23% of stools (154/673) while qPCR detected Shigella at diarrhea-associated quantities in 49% (329/673; P 94% sensitivity and specificity compared with the phenotypic susceptibility results for azithromycin and ampicillin. Performance for trimethoprim-sulfamethoxazole susceptibility was less robust and assessment of ciprofloxacin was limited because most isolates were resistant. Detection of AMR genes in direct stool generally yielded low specificities for predicting the resistance of the paired isolate, whereas the sensitivity and negative predictive values for predicting susceptibility were often higher. For example, detection of ermB or mphA in stool yielded a specificity of 56% but a sensitivity of 91% and negative predictive value of 91% versus the paired isolate. Patients that received azithromycin prior to presentation were universally culture negative (0/112), however qPCR still detected Shigella at diarrhea-associated quantities in 34/112 (30%). In sum, molecular diagnostics on direct stool greatly increases diagnostic yield for Shigella, including in the setting of prior antibiotics. Molecular detection of drug resistance genes in direct stool had low specificity for confirming resistance but could potentially "rule out" macrolide resistance.