Background and Aims: Diarrhoea has a significant health impact and requires accurate diagnostic approaches, despite the limitations of many existing methods. This review examines various molecular techniques aimed at facilitating the rapid diagnosis of diarrhoeal diseases caused by bacterial, viral and parasitic pathogens.

Methods: A comprehensive systematic literature review was conducted using six prestigious databases, including WOS, Scopus, Science Direct, Embase, PubMed and LILACS Plus. This rigorous approach allowed the synthesis of reviews on molecular diagnostic techniques for infectious diarrhoeal diseases.

Results: This research began with a careful systematic literature review of 2,760 scientific papers published in the last ten years (2014-2024), culminating in the inclusion of 18 studies following rigorous screening and eligibility criteria. In particular, multiplex polymerase chain reaction (PCR), isolation of genomic DNA from stool samples and the Luminex xTAG Gastrointestinal Pathogen Assay emerged as the predominant molecular methods. These techniques demonstrated remarkable consistency in sensitivity, specificity and rapid diagnostic capability, particularly in the context of acute infectious diarrhoeal diseases. They also demonstrated the ability to simultaneously detect and identify multiple pathogens, including bacteria, viruses and parasites.

Conclusions: The use of multiplex real-time PCR assays has not only improved the detection rates of enteropathogens, but has also revealed previously unrecognised gaps in the diagnosis of infectious diarrhoea. Our study highlights the importance of using molecular methods for comprehensive diagnosis in terms of sensitivity, specificity and rapid diagnostic capability. This will provide healthcare professionals with timely and accurate diagnostic data, enabling more effective treatment strategies for this public health problem.