Dataset Information

Development of LAMP assays for the molecular detection of taeniid infection in canine in Tibetan rural area.

ABSTRACT: For field-identification of taeniid cestodes in canine animals in Tibetan area, loop-mediated isothermal amplification (LAMP) assays for Echinococcus multilocularis, E. shiquicus, Taenia hydatigena, T. multiceps, T. pisiformis and T. crassiceps were developed and evaluated along with the reported assay for E. granulosus. The LAMP assays showed specific reaction with their corresponding target species DNA with the detection limit of 1 to 10 pg. Moreover, the assays for E. granulosus, E. multilocularis, T. hydatigena and T. multiceps could detect DNA extracted from 3 or more eggs of their corresponding target species. Then, the LAMP assays were applied on samples containing 3 to 35 taeniid eggs obtained from 61 field-collected canine feces in Qinghai, and the result was compared with a reported multiplex PCR and sequence analysis. The LAMP assays and the PCR detected single species DNA of E. granulosus, E. shiquicus, T. hydatigena and T. multiceps in 5, 2, 44 and 2 samples, respectively. In the rest 8 samples, DNA of both E. granulosus and T. hydatigena were detected by the PCR but the LAMP assays detected those DNAs in 2 samples and only T. hydatigena DNA in 6 samples. It was assumed that less than 3 E. granulosus eggs were mixed in the samples although the samples contained 21 to 27 eggs in total. In conclusion, the LAMP assays were less sensitive than the multiplex PCR, but would have adequate sensitivity for field use in Tibetan area.

SUBMITTER: Feng K

PROVIDER: S-EPMC5745177 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Development of LAMP assays for the molecular detection of taeniid infection in canine in Tibetan rural area.

Feng Kai K Li Wei W Guo Zhihong Z Duo Hong H Fu Yong Y Shen Xiuying X Tie Cheng C E Rijie R Xiao Changqin C Luo Yanhong Y Qi Guo G Ni Ma M Ma Qingmei Q Yamazaki Wataru W Yoshida Ayako A Horii Yoichiro Y Yagi Kinpei K Nonaka Nariaki N

The Journal of veterinary medical science 20171023 12

For field-identification of taeniid cestodes in canine animals in Tibetan area, loop-mediated isothermal amplification (LAMP) assays for Echinococcus multilocularis, E. shiquicus, Taenia hydatigena, T. multiceps, T. pisiformis and T. crassiceps were developed and evaluated along with the reported assay for E. granulosus. The LAMP assays showed specific reaction with their corresponding target species DNA with the detection limit of 1 to 10 pg. Moreover, the assays for E. granulosus, E. multilocu ...[more]

PMID: 29057765

Similar Datasets

Project description:Loop-mediated isothermal amplification (LAMP) is an alternative to PCR that is faster and requires fewer resources. Here, we describe two LAMP assays for the detection of human adenoviruses in the feces of children with acute intestinal infections. We designed сolorimetric LAMP (c-LAMP) and real-time LAMP (f-LAMP) with fluorescent probes to detect the DNA of the adenovirus F human adenovirus 40/41 (hAdV40/41) hexon gene. The detection limit of both developed methods was 103 copies/mL, which is comparable to the sensitivity of PCR. The specificities of both c-LAMP and f-LAMP were high, with no false-positive results for clinical samples that do not contain adenovirus F, when testing other viruses and microorganisms. Comparative tests of PCR and LAMP on clinical samples from patients with acute gastroenteritis were carried out. For all samples with a PCR threshold cycle (CT) of up to 36, the PCR and LAMP results completely coincided; however, at low viral loads, the diagnostic sensitivity of LAMP, especially c-LAMP with colorimetric detection, was inferior to that of PCR. The combination of LAMP with modern methods of nucleic acid extraction, both in manual and automatic modes, can reduce the time for a complete study, including extraction of nucleic acid material and amplification, to 60 min. IMPORTANCE In April 2022, several cases of acute hepatitis of unknown origin were reported in children from 12 countries. In many cases, enteric adenovirus or SARS-CoV-2 and adenovirus coinfection were detected. It is known that human adenoviruses can cause different infections of varying severity, from asymptomatic to severe cases with lethal outcomes. There is a need to increase the diagnostic capabilities of clinical laboratories to identify such an underestimated pathogen as adenovirus. Although PCR remains the gold standard for pathogen detection, this method requires specialized equipment and has a long turnaround time to process samples. Previously, LAMP assays for the detection of human adenovirus have been based on measuring the turbidity, the fluorescence of intercalated dyes, or electrophoretic separation. Herein, we present LAMP-based assays with colorimetric or fluorescent detection and perform a detailed assessment of their sensitivity, specificity, and diagnostic performance.

Project description:BackgroundMalaria in Cameroon is due to infections by Plasmodium falciparum and, to a lesser extent, Plasmodium malariae and Plasmodium ovale, but rarely Plasmodium vivax. A recent report suggested "Plasmodium vivax-like" infections around the study area that remained unconfirmed. Therefore, molecular and antigenic typing was used to investigate the prevalence of P. vivax and Duffy in asymptomatic adults resident in Bolifamba.MethodsA cross-sectional study was conducted from July 2008 to October 2009. The status of all parasite species was determined by nested PCR in 269 blood samples collected. The P. falciparum and P. vivax anti-MSP/CSP antibody status of each subject was also determined qualitatively by a rapid card assay. Parasite DNA was extracted from a sample infected with three parasite species, purified and sequenced. The Duffy antigen status of 12 subjects infected with P. vivax was also determined by sequencing. In silico web-based tools were used to analyse sequence data for similarities and matches to reference sequences in public DNA databases.ResultsThe overall malaria parasite prevalence in 269 individuals was 32.3% (87) as determined by PCR. Remarkably, 14.9% (13/87) of infections were caused either exclusively or concomitantly by P. vivax, established both by PCR and microscopic examination of blood smears, in individuals both positive (50%, 6/12) and negative (50%, 6/12) for the Duffy receptor. A triple infection by P. falciparum, P. vivax and P. malariae, was detected in one infected individual. Anti-MSP/CSP antibodies were detected in 72.1% (194/269) of samples, indicating high and continuous exposure to infection through mosquito bites.DiscussionThese data provide the first molecular evidence of P. vivax in Duffy positive and negative Cameroonians and suggest that there may be a significant prevalence of P. vivax infection than expected in the study area. Whether the P. vivax cases were imported or due to expansion of a founder effect was not investigated. Notwithstanding, the presence of P. vivax may complicate control efforts if these parasites become hypnozoitic or latent as the liver stage.ConclusionsThese data strongly suggest that P. vivax is endemic to the south-west region of Cameroon and should be taken into account when designing malaria control strategies.

Project description:Tibetan foxes (Vulpes ferrilata) have been confirmed as the main wild definitive hosts in echinococcosis transmission in the eastern Tibetan Plateau. However, little information is available about the epidemiology in wildlife from the perspective of the Taeniidae family, which is essential knowledge in understanding the epidemiology and phylogeography of cestode species in the Tibetan plateau. Therefore, in this study, we used copro-PCR techniques, by amplifying nad1 and cox1 gene fragments, to detect the taeniid species from Tibetan fox feces collected in Shiqu County, (Sichuan Province, China), eastern Tibetan Plateau. Phylogenetic relationships between amplified sequences and existed Taenia species genotypes were evaluated. Then, the maximum prevalence (positive PCR results from at least one primer pair) and the conservative prevalence (positive PCR results from at least two primer pairs) were calculated. Thirty-six Tibetan fox feces were analyzed. Echinococcus multilocularis (conservative prevalence ± 95% CI: 22.2% ± 13.6%; maximum prevalence ± 95% CI: 33.3% ± 15.4%) and E. shiquicus (2.8 ± 5.4%; 8.3 ± 9.0%) was detected. Meanwhile, DNA fragments of T. polyacantha were detected with high similarity to NCBI sequences (cox1, 94.0%) and to the larva sample DNA sequenced in this study (93.4%), and were supported by phylogenetic analysis. Thus, T. polyacantha might infect Tibetan foxes (5.6% ± 7.5%, 11.1% ± 10.3%). Our limited findings in the epidemiology of parasitic Taenia species suggest that sylvatic transmission cycles for a more species-rich Taeniid community must be established between wild canids and small mammals than just for the two Echinococcus species. Besides, discrepancies in different primer pairs in detecting the taeniid species were evaluated. The sensitivity of some widely used universal primer pairs was poor in detecting Taenia species from canid copro-DNA samples. It is still challenging to the development of effective taeniid species-specific molecular markers especially for non-zoonotic species.

Dataset Information

Development of LAMP assays for the molecular detection of taeniid infection in canine in Tibetan rural area.

Publications

Development of LAMP assays for the molecular detection of taeniid infection in canine in Tibetan rural area.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets