Molecular typing of dengue viruses circulating in Assam, India during 2016-2017.

BACKGROUND & OBJECTIVES: The global incidence of dengue has grown dramatically in recent decades and Assam, India has witnessed several outbreaks of dengue since 2015. Although during post-monsoon months (September to December), most cases of dengue in Assam are recorded but incidence of dengue in Assam has been slowly changing from being endemic to being hyper endemic. Therefore, this study was carried out to determine the serotypes and genotypes of dengue virus prevalent in Assam during the period of 2016-2017. METHODS: This is a prospective study conducted for a period of two years from 2016 to 2017. Department of Microbiology, Gauhati Medical College and Hospital (GMCH) had received a total of ~12000 and ~9000 sera sample during 2016 and 2017 respectively for confirmation of clinically suspected dengue cases. For confirmation, dengue NS1 antigen and IgM antibody ELISA tests were performed. Multiplex RT-PCR was performed for serotyping of dengue viruses and representative samples found positive in PCR were sequenced to determine the genotypes of circulating dengue virus serotypes. RESULTS: In the year 2016, 6157 sera samples and in 2017, 3386 sera samples were found positive in ELISA test. A total of 157 dengue positive sera samples representing 17 districts of Assam were further tested by multiplex RT-PCR for serotyping of the virus. In PCR, out of 157, 107 samples (68.15%) were found positive for the presence of dengue virus genome. Out of 107, 74 samples (69.15%) were positive for dengue virus serotype-1 (DENV-1), 32 samples (29.90%) for dengue virus serotype-2 (DENV-2) and one sample (0.93%) positive for dengue virus serotype-3 (DENV-3). Out of 107 PCR positive samples, 25 samples were sequenced to identify their genotypes. Phylogenetic analysis of sequenced dengue viruses revealed that all the seven DENV-1 strains were genotype V, 17 DENV-2 strains were genotype IV (Cosmopolitan genotype) and one DENV-3 strain was genotype III. INTERPRETATION & CONCLUSION: These findings improve our knowledge of circulating dengue virus serotypes in Assam. Co-circulation of three serotypes of dengue virus highlights the need for establishment of active dengue surveillance. The genotypic data of our findings will be helpful for future dengue molecular epidemiology studies and to control the disease in the region.

Design of a 3D printed smartphone microscopic system with enhanced imaging ability for biomedical applications.

Portable, low-cost smartphone platform microscopic systems have emerged as a potential tool for imaging of various micron and submicron scale particles in recent years (Ozcan; Pirnstill and Coté; Breslauer et al.; Zhu et al.). In most of the reported works, it involves either the use of sophisticated optical set-ups along with a high-end computational tool for postprocessing of the captured images, or it requires a high-end configured smartphone to obtain enhanced imaging of the sample. Present work reports the working of a low-cost, field-portable 520× optical microscope using a smartphone. The proposed smartphone microscopic system has been designed by attaching a 3D printed compact optical set-up to the rear camera of a regular smartphone. By using cloud-based services, an image processing algorithm has been developed which can be accessed anytime through a mobile broadband network. Using this facility, the quality of the captured images can be further enhanced, thus obviating the need for dedicated computational tools for postprocessing of the images. With the designed microscopic system, an optical resolution ∼2 µm has been obtained. Upon postprocessing, the resolution of the captured images can be improved further. It is envisioned that with properly designed optical set-up in 3D printer and by developing an image processing application in the cloud, it is possible to obtain a low-cost, user-friendly, field-portable optical microscope on a regular smartphone that performs at par with that of a laboratory-grade microscope. LAY DESCRIPTION: With the ever-improving features both in hardware and software part, smartphone becomes ubiquitous in the modern civilised society with approximately 8.1 billion cell phone users across the world, and ∼40% of them can be considered as smartphones. This technology is undoubtedly the leading technology of the 21st century. Very recently, various researchers across the globe have utilised different sensing components embedded in the smartphone to convert it into a field-portable low-cost and user-friendly tool which can be used for different sensing and imaging purposes. By using simple optical components such as lens, pinhole, diffuser etc. and the camera of the smartphone, various groups have converted the phone into a microscopic imaging system. Again, by removing the camera lenses of the phone, holography images of microscopic particles by directly casting its shadows on the CMOS sensor on the phone has been demonstrated. The holographic images have subsequently been processed using the dedicated computational tool, and the original photos of the samples can be obtained. All the reported smartphone-based microscopic systems either suffer from relatively low field-of-view (FOV), resolution or it needs a high computational platform. Present work, demonstrate an alternative approach by which a reasonably good resolution (<2 µm) along with high optical magnification (520×) and a large FOV (150 µm) has been obtained on a regular smartphone. For postprocessing of the captured images an image processing algorithm has been developed in the cloud and the same can be accessed by the smartphone application, obviating the need of dedicated computational tool and a high-end configured smartphone for the proposed microscope. For the development of the proposed microscopic system, a simple optical set-up has been fabricated in a 3D printer. The set-up houses all the required optical components and the sample specimen with the 3D-printed XY stage, and it can be attached easily to the rear camera of the smartphone. Using the proposed microscopic system, enhanced imaging of USAF target and red blood cells have been successfully demonstrated. With the readily available optical components and a regular smartphone, the net cost involvement is significantly low (less than $250, including the smartphone). We envisioned that the designed system could be utilised for point-of-care diagnosis in resource-poor settings where access to the laboratory facilities is very limited.