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Tropical Medicine and Infectious Disease Jun 2024Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic...
Rapid Discriminative Identification of the Two Predominant Echinococcus Species from Canine Fecal Samples in the Tibetan Region of China by Loop-Mediated Isothermal Amplification-Lateral Flow Dipstick Assay.
Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic subtypes are cystic echinococcosis (CE) and alveolar echinococcosis (AE), which result from infestation by and , respectively. A prominent epidemic of echinococcosis greatly affects the Tibet Autonomous Region (TAR) in China. A new technique called the loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) test is introduced in this research to differentiate between and using their repetitive genetic sequences. The test is characterized by its portable nature, simple operation, quick result production, high sensitivity, and low susceptibility to aerosol contamination. The LAMP-LFD method demonstrated an exceptional minimal detection limit, reaching levels as low as approximately 1 fg/μL (femtogram per microliter) of genomic DNA. The assay's specificity was assessed, and no cross-reactivity was seen. A total of 982 dog fecal samples were collected from 54 counties in the TAR region between July 2021 and June 2022. The established method underwent validation using a commercially available ELISA kit. The agreement rate between the LAMP-LFD and ELISA methods was 97.25%, with a sensitivity of 96.05% and a specificity of 97.35%. The assay described in this study improves specificity by using a double-labeled probe, and it reduces the risk of false-positive results caused by aerosol contamination through the use of a sealed device. This makes it a suitable choice for quickly and accurately identifying the two main types of in field settings.
PubMed: 38922048
DOI: 10.3390/tropicalmed9060136 -
Scientific Reports Jul 2023Lyme disease, one of the most common tickborne diseases, has been rapidly spreading in parallel with the expansion of the range of its tick vector. Better tick...
Lyme disease, one of the most common tickborne diseases, has been rapidly spreading in parallel with the expansion of the range of its tick vector. Better tick surveillance efforts are needed to accurately estimate disease risk and to guide public health and clinical management. We have developed two multiplex loop-mediated isothermal amplification (LAMP) reactions coupled with oligonucleotide strand displacement (OSD) probes to identify the tick host, Ixodes scapularis, and the Lyme disease pathogen, Borrelia burgdorferi, they carry. In each multiplex LAMP-OSD assay the co-presence of two target sequences is computed at the DNA level by linking the two corresponding amplicons and detecting the co-product on colorimetric lateral flow dipsticks. In tests with synthetic DNA, the co-presence of as few as four copies of input DNA could be detected, without producing spurious signals. Most importantly, though, the LAMP-OSD assay is amenable to being carried out directly with macerated tick samples, without any sample preparation. In such field conditions, assays performed robustly and demonstrated 97-100% sensitivity and 100% specificity with both field-collected and lab-raised artificially infected ticks. Such easy-to-use, arthropod and pathogen-specific assays would be well suited to field and near patient use without relying on complex instrumentation or infrastructure.
Topics: Animals; Humans; Ixodes; Borrelia; Colorimetry; Lyme Disease; Borrelia burgdorferi; Nucleic Acids
PubMed: 37454160
DOI: 10.1038/s41598-023-38452-8 -
RSC Advances Jan 2024A reversible acid-base probe, (NE, NE)-N, N-bis(()-3-(4-(dimethylamino)phenyl)allylidene)benzene-1,4-diamine (MM1), is introduced for the colorimetric visualization of...
A reversible acid-base probe, (NE, NE)-N, N-bis(()-3-(4-(dimethylamino)phenyl)allylidene)benzene-1,4-diamine (MM1), is introduced for the colorimetric visualization of acid-base equilibria in non-aqueous media. MM1 displays reversible acidochromic behavior, showing exciting colorimetric change varying from weak to strong acid. Also, we have fabricated a colorimetric paper strip-based test kit to visualize acid-base equilibria. A dipstick experiment has been demonstrated to visualize the acid-base equilibria in the gaseous state. This acid-base probe has also been employed to estimate the p values of several acidic compounds in a non-aqueous medium using overlapping indicator methods. Based on reversible acidochromic UV-visible absorption spectral and colorimetric behavior, we have constructed a reconfigurable dual input and dual output combinational logic circuit and set-reset memorized device employing acid and base as chemically encoded inputs and corresponding optical outputs. The current report evokes a new protocol for developing various reversible acidochromic probes and its implication for constructing opto-chemical molecular logic gates and estimating the acid dissociation constants of various acidic compounds in non-aqueous media.
PubMed: 38259994
DOI: 10.1039/d3ra04696f