Environmental detection of Burkholderia pseudomallei and associated melioidosis risk: a molecular detection and case-control cohort study

17 Jun 2025·
Sukritpong Pakdeerat
,
Chalita Chomkatekaew
,
Phumrapee Boonklang
Arin Wongprommoon
Arin Wongprommoon
,
Kesorn Angchagun
,
Yaowaret Dokket
,
Areeya Faosap
,
Gumphol Wongsuwan
,
Premjit Amornchai
,
Vanaporn Wuthiekanun
,
Jirameth Changklom
,
Suwatthiya Siriboon
,
Parinya Chamnan
,
Sharon J Peacock
,
Jukka Corander
,
Nicholas PJ Day
,
Nicholas R Thomson
,
Chayasith Uttamapinant
,
Somsakul Pop Wongpalee
,
Claire Chewapreecha
· 2 min read
Abstract
Environmental exposure to Burkholderia pseudomallei, the causative agent of melioidosis, remains poorly characterised due to the limited sensitivity of conventional detection methods. This hinders accurate risk mapping and delays public health responses. Here, we developed CRISPR-BEEPs – a sensitive, equipment-light CRISPR-based assay – that demonstrated substantially improved sensitivity (93.5% vs 19.4%) and specificity (100% vs 98.0%) compared to conventional culture-based plate inspection techniques. We applied CRISPR-BEEPs to water samples collected from both natural and piped sources across 15,118 km2 in northeast Thailand, including households of confirmed melioidosis patients and controls. B. pseudomallei was detected in 73.3% of groundwater samples, 32.4% of surface water, and 28.3% of piped water, with peak detection during the flood season. Importantly, the assay’s improved sensitivity enabled detection of a significant association between environmental B. pseudomallei detection within a 10 km radius of participants’ households and melioidosis risk (OR 2.74; 95% CI = 1.38–5.48) – an association undetectable by conventional methods. These findings highlight major gaps in water treatment and distribution infrastructure and demonstrate the value of high-resolution environmental diagnostics. Strengthening water sanitation and surveillance systems is essential for mitigating melioidosis transmission and addressing the broader burden of waterborne diseases in vulnerable settings.
Type
Publication
medRxiv

Lay summary

Melioidosis is a neglected tropical disease caused by the bacterium Burkholderia pseudomallei, which can be acquired from the environment through contact with soil or water. However, no previous studies established a statistically significant relationship between B. pseudomallei in the local environment and melioidosis cases, mostly because of low abundance of the bacterium and inconsistent sampling methods.

We recently developed a CRISPR-based method to detect B. pseudomallei and compared it with conventional microbial culture methods on how well it detects the bacterium in water samples from northeast Thailand. We found that our method is far more sensitive (able to pick up the bacterium in most samples with it) and equally specific (able to rule out the bacterium in most samples without it) compared to culture-based methods.

Additionally, we collected water samples, clinical information, and demographic information associated with people in the endemic area. We found exposure to B. pseudomallei in water sources, being an agricultural worker, and diabetes increased the risk of melioidosis.

My role

I performed a literature search which shows that molecular-based techniques (detecting DNA or RNA) had twice the test positivity rate as culture-based techniques (growing the bacterium on agar and finding colonies by eye).

I also processed and analysed the clinical data: I grouped the study participants’ occupation codes and produced a summary of the demographic characteristics of the population (ages, sexes, BMIs, blood glucose level, etc.).