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Diagnosis of gastrointestinal parasites has traditionally relied on stool microscopy, which has low diagnostic sensitivity and specificity. We have developed a novel, rapid, high-throughput quantitative multi-parallel real-time polymerase chain reaction (qPCR) platform. Species-specific primers/probes were used for eight common gastrointestinal parasite pathogens: Ascaris lumbricoides, Necator americanus, Ancylostoma duodenale, Giardia lamblia, Cryptosporidium spp., Entamoeba histolytica, Trichuris trichiura, and Strongyloides stercoralis. Stool samples from 400 13-month-old children in rural Ecuador were analyzed and the qPCR was compared with a standard direct wet mount slide for stool microscopy, as were 125 8-14-year-old children before and after anthelmintic treatment. The qPCR showed higher detection rates for all parasites compared with direct microscopy, Ascaris (7.0% versus 5.5%) and for Giardia (31.5% versus 5.8%). Using an enhanced DNA extraction method, we were able to detect T. trichiura DNA. These assays will be useful to refine treatment options for affected populations, ultimately leading to better health outcomes.

Multi Parallel - Multiple Accounts & App Clone is a free utility application for mobile devices developed by Winterfell Applab. It is a cloning tool that can help users create multiple instances of a single app on their smartphone. It supports various popular social media programs and games.

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Multi Parallel is essentially a utility to help you create multiple clones of your preferred application or game. Its usefulness will vary from user to user, but it is most commonly used with social media apps and MMO games where it is common for a person to have more than one account on a single device. With it, you will be able to manage multiple accounts and quickly switch between them with ease.

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KK may be an inadequate tool for stool-based surveillance in areas where hookworm or Strongyloides are common or where intensity of helminth infection is low after repeated rounds of chemotherapy. Because deworming programs need to distinguish between populations where parasitic infection is controlled and those where further treatment is required, multi-parallel qPCR (or similar high throughput molecular diagnostics) may provide new and important diagnostic information.

Although they are costly, these techniques are already used in centralized laboratories in many low- and middle-income countries. Molecular diagnostics for a wide range of infectious disease agents can improve sensitivity and specificity of measurements, allow for more accurate quantitation, increase throughput, require fewer trained personnel and offer insight into the relationships among multiple pathogens. From a practical perspective, qPCR assays may be particularly useful to monitor the intensity of STH (and other parasites) in areas nearing transmission elimination after repeated rounds of chemotherapy. They provide a more precise way to examine trends in low-intensity infection settings, determine if reinfection occurs post-treatment, and assess interruption of transmission in a defined setting.

This study was designed to measure the benefit of a novel molecular diagnostic tool relative to the traditional KK egg-counting technique commonly used in deworming impact evaluations. Our primary aim was to assess whether, and to what degree, high-throughput, multi-parallel qPCR increased sensitivity and diagnostic breadth compared to traditional stool-based parasitic diagnosis. A second aim was to assess whether qPCR could be used to measure the intensity of infection. As A. lumbricoides and N. americanus were the only STH present in the study setting beyond trace levels, this study focuses on these species.

Three months post-treatment, a follow-up parasitological survey and worm expulsion study were carried out using the same procedure as described above. Some people were present only at baseline or only at follow-up. There were 796 people with complete KK and qPCR records at baseline; 633 of these people also had complete records at follow-up. The total number of stools that were tested both by KK and by qPCR is 1884. This sample includes people who were present at only one time-point, stool samples from the efficacy time-point, and multiple stools from some individuals.

Based on the present study, the prevalence of STH is likely to be underestimated by current mapping efforts in many low endemicity settings. As an alternative, qPCR can be used to more accurately assess changes in infection prevalence and intensity following MDA. This is particularly important as periodic MDA continues to bring down the prevalence of STH. Although qPCR requires transporting samples to reference laboratories where they can be processed, the use of such molecular techniques allows for more sensitive measurements of infection prevalence and intensity. Multi-parallel qPCR for STH can be more easily undertaken in resource-limited settings than can multiplex qPCR, as more standard (and accessible) instrumentation can be used [32]. KK still serves an important purpose for mapping areas of presumed high STH and Schistosoma spp. prevalence, but thorough examinations of the comparative costs of qPCR and KK under field conditions are necessary. Though monitoring and evaluation teams are currently asking whether they can afford to use qPCR for STH surveillance, they may soon be asking whether they can afford to lose so much information by putting off building their capacity in modern molecular methods.

Small-scale, multi-parallel bioreactors provide more process information for culture comparison in less time. Multiple experiments can assess different clones or cell lines, as well as the effects of pH, temperature, feeding, media, gassing rates and inoculation densities.

Ambr multi-parallel bioreactors can handle your upstream process steps, from clone or cell line selection to process optimization, ensuring scalability because process data translates well into the 2L Univessel SU and larger Biostat STR systems.

The qPCR has been useful in the diagnosis of pathogenic microbes causing tissue infections, including amebic liver abscess, and has also been shown to have a high degree of sensitivity and specificity in the detection of enteric pathogens.2,3,9 As with other real-time PCR systems, to broaden the spectrum of enteric pathogen detection, a simple to execute, multi-parallel qPCR approach was developed that enables detection of an unlimited number of a parasite species provided that sequence information is available.

These assays were validated by using genomic DNA spiked into known negative stool and/or stools from patients known to be parasite positive by standard stool examination (performed by Clinical Center Microbiology Laboratory, National Institutes of Health, Bethesda, MD) by using standard techniques (e.g., direct microscopy, modified acid-fast staining for Cryptosporidium, and agar plate migration for S. stercoralis).6 Validation using similar techniques had been performed for all parasites except T. trichiura.2,3 In addition, stools cryopreserved in the United States, Mali, or India (as part of other studies) that had multiple negative results for stool examinations for helminth eggs and larvae were tested by using qPCR. All samples showed negative results.

Although direct observation of a stool sample in a wet smear, Kato-Katz slides, and stool concentration methods have been used for diagnosing diarrheal illness and for evaluation of eosinophilia, the sensitivity and specificities of microscopy depend on the type of organism, its life cycle, and the pattern of egg or larval shedding. Although qPCR also relies on these factors, the ability to detect attograms of parasitic DNA overcomes many of these potential obstacles because of higher sensitivity. In the present study, we compared our qPCR to direct smear microscopy primarily to normalize the conditions between the two tests (Table 2). Although the Kato-Katz/fecal concentration method generally has a higher sensitivity than direct microscopy, and for A. lumbricoides and T. trichiura is semi-quantitative, it is more time-consuming and relies on collection of multiple stool samples to maximize sensitivity. Antigen detection assays with improved parasite detection ability are available (e.g., for G. lamblia and E. histolytica), but these assays are costly and not readily available in resource-limited areas.7,13 e24fc04721