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Intestinal diarrheagenic polyparasitic infections are among the major public health concerns in developing countries. Here we examined stool specimens by microscopy, DNA dot blot and polymerase chain reaction (PCR) to evaluate the co-infection of four principal protozoans among amoebic dysentery cases from Northeast Indian population. The multiplex PCR confirmed Entamoeba histolytica (8·1%), Entamoeba dispar (4·8%) and mixed infection of both the parasites (3·4%) in 68 of 356 stool specimens that were positive in microscopy and/or HMe probe based DNA dot blot screening. The prevailing parasite that co-exists with E. histolytica was Giardia duodenalis (34·1%), followed by Enterocytozoon bieneusi (22·0%), Cryptosporidium parvum (14·6%) and Cyclospora cayetanensis (7·3%, P = 0·017). Symptomatic participants (odds ratio (OR) = 4·07; 95% confidence interval (CI) = 1·06, 15·68; P = 0·041), monsoon season (OR = 7·47; 95% CI = 1·40, 39·84; P = 0·046) and participants with family history of parasitic infection (OR = 4·50; 95% CI = 1·16, 17·51; P = 0·030) have significant association with overall co-infection rate. According to molecular consensus, comprehensive microscopy yielded 3·4% (12/356) false-negative and 7·6% (27/356) false-positive outcome, suggesting an improved broad-spectrum PCR-based diagnostic is required to scale down the poor sensitivity and specificity as well as implementation of integrated control strategy. [ABSTRACT FROM PUBLISHER]

Paper-based DNA biosensors are powerful tools in point-of-care diagnostics since they are affordable, portable, user-friendly, rapid and robust. However, their sensitivity is not always as high as required to enable DNA quantification. To improve the response of standard dot blots, we have applied a new enhancement strategy that increases the sensitivity of assays based on the use of biotinylated silica-nanoparticles (biotin-Si-NPs). After immobilization of a genomic Campylobacter DNA onto a paper membrane, and addition of a biotinylated-DNA detection probe, hybridization was evidenced using streptavidin-conjugated to horseradish peroxidase (HRP) in the presence of luminol and H 2 O 2. Replacement of the single biotin by the biotin-Si-NPs boosted on average a 30 fold chemiluminescent read-out of the biosensor. Characterization of biotin-Si-NPs onto a paper with immobilized DNA was done using a scanning electron microscope. A limit of detection of 3 pg/μL of DNA, similar to the available qPCR kits, is achieved, but it is cheaper, easier and avoids inhibition of DNA polymerase by molecules from the food matrices. We demonstrated that the new dot blot coupled to biotin-Si-NPs successfully detected Campylobacter from naturally contaminated chicken meat, without needing a PCR step. Hence, such an enhanced dot blot paves the path to the development of a portable and multiplex paper based platform for point-of-care screening of chicken carcasses for Campylobacter. Image 1 • Enhanced chemiluminescent signal on DNA dot blot. • Single extra step involving biotinylated Si-nanoparticles boosts the signal 30 times. • Paper-based detection of bacterial DNA without pre-amplification. • Useful technology to monitor Campylobacter in naturally contaminated chicken meat. [ABSTRACT FROM AUTHOR]