In 2013 a new reassortant avian influenza A H7N9 virus emerged in China, causing human infection with high mortality. An accurate and timely diagnosis is crucial for controlling the outbreaks of the disease. We therefore propose a simple strategy for rapidly and sensitively detecting the H7N9 virus using an intensity-modulated surface plasmon resonance (IM-SPR) biosensor integrated with a new generated monoclonal antibody. The novel antibody exhibits significant specificity to recognize H7N9 virus compared with other clinical human influenza isolates (p < 0.01). Experimentally, the detection limit of the proposed approach for H7N9 virus detection is estimated to be 144 copies/mL, which is a 20-fold increase in sensitivity compared with homemade target-captured ELISA using the identical antibody. For the measurement of mimic clinical specimens containing the H7N9 virus mixed with nasal mucosa from flu-like syndrome patients, the detection limit is calculated to be 402 copies/mL, which is better than conven...

Simple Strategy for Rapid and Sensitive Detection of Avian Influenza A H7N9 Virus Based on Intensity-Modulated SPR Biosensor and New Generated Antibody

Infectious diseases are clinically relevant, transmissible illnesses caused by microorganisms, such as bacteria, viruses, fungi, and parasites. According to a recent report, between 1980 and 2014 more than four million people in the USA have died from various infectious diseases.1 Accurate and timely detection of infection and identification of the causative pathogen are crucial in disease prevention, treatment and monitoring.2,3 Although reliable methods of pathogen detection are well and long established, many of such detection and diagnostics techniques are still limited in use to clinical laboratories due to the need for trained and skilled personnel, and sophisticated diagnostic instruments. Accordingly, there is a pressing need for simple, affordable, and easy-to-use diagnostic tools for the specific detection of pathogens at the point of care, e.g., doctors' offices, clinics, infirmaries, and particularly in resource-limited areas where medical infrastructure is lacking.

Interfacing Pathogen Detection with Smartphones for Point-of-Care Applications

India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

/pdf/loop-mediated-isothermal-amplification-an-innovative-gene-2hl5486a5y.pdf

Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases.

https://dspace.stir.ac.uk/bitstream/1893/21852/1/El%20Wahed%20et%20al_JCV_2015.pdf

Diagnostics-in-a-Suitcase: Development of a portable and rapid assay for the detection of the emerging avian influenza A (H7N9) virus.

Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA), is a major bacterial pathogen associated with nosocomial and community-acquired S. aureus infections all over the world. A rapid detection assay for staphylococcal gene of nuc and mecA is needed. In this study, a rapid identification assay based on the loop-mediated isothermal amplification (LAMP) method was established. PCR and LAMP assays were used to detect Staphylococcus aureus and other related species for nuc and mecA. With optimization of the primers and reaction temperature, the LAMP successfully amplified the genes under isothermal conditions at 62 °C within 60 min, of which the results were identical with those of the conventional PCR methods. The detection limits of the LAMP for nuc and mecA were 1.47 and 14.7 pg/μl DNA per tube, respectively, by naked eye inspections, while the detection limits of the PCR for nuc and mecA were 14.7 pg/μl and 147 pg/μl DNA, respectively. Finally, The LAMP method was then applied to clinical blood plaque samples. The LAMP and PCR demonstrated identical results for the plaque samples with the culture assay. Together, the LAMP offers an alternative detection assay for nuc and mecA with a great advantage of the sensitivity and rapidity.

Rapid Detection of Staphylococcus Aureus by Loop-Mediated Isothermal Amplification

An epidemic of human H7N9 influenza virus infection recently emerged in China whose clinical features include high mortality and which has also resulted in serious economic loss. The novel reassortant avian-origin influenza A (H7N9) virus which was the causative agent of this epidemic raised the possibility of triggering a large-scale influenza pandemic worldwide. It seemed likely that fast molecular detection assays specific for this virus would be in great demand. Here, we report a one-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of the hemagglutinin (HA) and neuraminidase (NA) genes of H7N9 virus, the minimum detection limit of which was evaluated using in vitro RNA transcription templates. In total, 135 samples from clinical specimens (from either patients or poultry) were tested using this method in comparison with the real-time PCR recommended by the World Health Organization (WHO). Our results showed that (i) RT-LAMP-based trials can be completed in approximately 12 to 23 min and (ii) the detection limit for the H7 gene is around 10 copies per reaction, similar to that of the real-time PCR, whereas the detection limit for its counterpart the N9 gene is 5 copies per reaction, a 100-fold-higher sensitivity than the WHO-recommended method. Indeed, this excellent performance of our method was also validated by the results for a series of clinical specimens. Therefore, we believe that the simple, fast, and sensitive method of RT-LAMP might be widely applied for detection of H7N9 infections and may play a role in prevention of an influenza pandemic.

https://jcm.asm.org/content/jcm/51/11/3760.full.pdf

Rapid and Sensitive Detection of H7N9 Avian Influenza Virus by Use of Reverse Transcription–Loop-Mediated Isothermal Amplification

The invention relates to a primer, a kit and a detection method for detecting an avian influenza H7N9 virus by using an RT-LAMP method The primer comprises an H7 primer group and an N9 primer group and can effectively detect the avian influenza H7N9 virus The kit comprises the above-mentioned primer and can be used for detecting the avian influenza H7N9 virus The detection method employs the above-mentioned kit and can intuitively determine results according to the color of a reaction solution, being rapid and accurate According to the invention, the results are reliable; cost is low; the characteristics of easiness, rapidness and sensitivity are obtained; the primer, the kit and the detection method are especially suitable for on-site detection and applicable to most hospitals and laboratories of epidemic prevention departments lacking real-time fluorescence quantification PCR instruments in China

Primer, kit and detection method for detecting avian influenza H7N9 virus by using RT-LAMP method

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Rapid and Sensitive Detection of H7N9 Avian Influenza Virus by Use of Reverse Transcription–Loop-Mediated Isothermal Amplification

Primer, kit and detection method for detecting avian influenza H7N9 virus by using RT-LAMP method