Tuberculosis, caused by Mycobacterium tuberculosis, remains one of the most enduring public health challenges worldwide, particularly in low- and middle-income countries [1]. Despite considerable progress in TB control over the past decades, the World Health Organization (WHO) estimates that nearly a quarter of the world's population is infected with TB, underscoring the urgent need for effective diagnostic and treatment strategies [2,3]. The advent of rapid and accurate diagnostic tools is pivotal for early detection and management of TB, which is critical in curtailing its transmission and improving patient outcomes [4,5].

The diagnostic landscape of TB has significantly evolved with the introduction of molecular technologies that offer substantial improvements over conventional microbiological methods such as sputum smear microscopy and culture [6]. Among these, the GeneXpert MTB/RIF assay (Cepheid, Sunnyvale, CA, USA), a nucleic acid amplification test (NAAT), has been a revolutionary advancement [7,8]. Endorsed by the WHO as a frontline test for TB and rifampicin resistance detection, GeneXpert MTB/RIF combines the detection of TB DNA with the simultaneous identification of rifampicin resistance mutations, offering results in under two hours [9,10]. This rapid turnaround time ensures early initiation of appropriate treatment regimens, which is crucial for patient prognosis and preventing the transmission of drug-resistant TB strains [11].

Parallelly, CRISPR-based diagnostics have surfaced as a promising innovation in the field of infectious diseases, including TB. Leveraging the precision of the CRISPR/Cas system for genome editing, these diagnostics aim to detect specific genetic sequences of the TB pathogen, including drug-resistance mutations [12,13]. The specificity and adaptability of CRISPR technologies grant them the potential to not only provide accurate TB diagnoses but also facilitate personalized treatment strategies by identifying the most effective drug regimens for each patient's strain of TB [14,15]. Such capabilities position CRISPR-based diagnostics as a significant stride towards realizing the goals of precision medicine in TB management.

Moreover, Innowave DX MTB/RIF assays, employing various molecular techniques to detect TB DNA in clinical specimens, have been integral in the TB diagnostic arsenal [16]. Despite being overshadowed by the more advanced GeneXpert MTB/RIF and emerging CRISPR-based diagnostics in terms of specificity and sensitivity, Innowave DX MTB/RIF assays play a crucial complementary role, especially in settings where access to sophisticated technologies is limited [17].

The diverse patient populations affected by TB present unique diagnostic challenges, influenced by demographic factors (such as age and gender), as well as the presence of comorbidities like diabetes and hypertension which are known to affect the course and diagnosis of TB [18,19]. The interaction between TB and these comorbid conditions can complicate diagnosis, potentially leading to delayed treatment and adverse patient outcomes [20]. Furthermore, age-related physiological changes and gender-specific biological and social determinants may influence the performance of diagnostic tests [21]. Therefore, evaluating the diagnostic efficacy of these advanced technologies across different TB patient populations is paramount in ensuring their adaptability and utility in varied demographic and clinical settings.

Given this context, our study aimed to perform a comprehensive evaluation of the diagnostic efficacy of CRISPR-based TB diagnostics, GeneXpert MTB/RIF, and Innowave DX MTB/RIF across diverse TB patient populations. By elucidating the strengths and limitations of these diagnostic modalities in relation to demographic variables and comorbid conditions, we aspire to contribute valuable insights towards optimizing TB diagnosis and management. This effort aligns with global health objectives to curb the spread of TB and effectively manage the disease, ultimately propelling us closer to the goal of eradicating TB.