Abstract
BackgroundThe diagnosis of filariasis traditionally relies on the detection of circulating microfilariae (mf) using Giemsa-stained thick blood smears. This approach has several limitations. We developed a semi-automated microfluidic device to improve and simplify the detection of filarial nematodes.MethodsThe efficiency and repeatability of the microfluidic device was evaluated. Human EDTA blood samples were ‘spiked’ with B. malayi mf at high, moderate, and low levels, and subsequently tested 10 times. The device was also used for a field survey of feline filariasis in 383 domesticated cats in an area of Narathiwat Province, Thailand, the endemic area of Brugia malayi infection.ResultsIn the control blood arbitrarily spiked with mf, the high level, moderate level and low level mf-positive controls yielded coefficient variation (CV) values of 4.44, 4.16 and 4.66%, respectively, at the optimized flow rate of 6 µl/min. During the field survey of feline filariasis in Narathiwat Province, the device detected mf in the blood of 34 of 383 cats (8.9%) whereas mf were detected in 28 (7.3%) cats using the blood smear test. Genomic DNA was extracted from mf trapped in the device after which high-resolution melting (HRM) real-time PCR assay was carried out, which enabled the simultaneous diagnosis of filarial species. Among the 34 mf-positive samples, 12 were identified as B. malayi, 15 as Dirofilaria immitis and 7 as| D. repens.ConclusionsWe developed a semi-automated microfluidic device to detect mf of filarial parasites that could be used to diagnose lymphatic filariasis in human populations. This novel device facilitates rapid, higher-throughput detection and identification of infection with filariae in blood samples.
Highlights
The diagnosis of filariasis traditionally relies on the detection of circulating microfilariae using Giemsa-stained thick blood smears
We report a novel, semi-automated microfluidic device that integrates real-time and high resolution melting polymerase chain reaction (PCR) for the simultaneous detection and differentiation of species of filariae, in the blood of domesticated cats naturally infected with species of Brugia and Dirofilaria
Study samples To study the efficiency and reproducibility of the semiautomated microfluidic device, three groups of samples were prepared by spiking microfilariae of B. malayi into EDTA blood obtained from a healthy subject
Summary
The diagnosis of filariasis traditionally relies on the detection of circulating microfilariae (mf ) using Giemsa-stained thick blood smears. We developed a semi-automated microfluidic device to improve and simplify the detection of filarial nematodes. The microfluidic approach has been applied for the detection of diverse pathogens including blood stages of malarial parasites [6,7,8,9]. Microfluidicbased platforms offer advantages in comparison to thick blood smear staining technique, including speed, cost, portability, high throughput and automation [9]. Microfluidic technologies have been used in research on other parasitic nematodes, including the deployment of a chip device that integrates microfluidics and electrophysiology to provide real-time records of the nano-scale electrical signals emitted by nematode muscles and neurons [11], and a microfluidic electropharyngeograms to screen anthelmintic candidates and investigate feeding behaviors by nematodes for the discovery of novel anthelmintics [12]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
