scholarly journals Efficiency of the Stool-PCR Test Targeting NADH Dehydrogenase (Nad) Subunits for Detection of Opisthorchis viverrini Eggs

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Wansika Phadungsil ◽  
Supaporn Pumpa ◽  
Kridsada Sirisabhabhorn ◽  
Amornrat Geadkaew-Krenc ◽  
Rudi Grams ◽  
...  
Keyword(s):  
Mitochondrial Gene ◽  
Parasitic Infection ◽  
Pcr Amplification ◽  
Detection Limits ◽  
Cross Reactivity ◽  
Opisthorchis Viverrini ◽  
Stool Examination ◽  
Special Equipment ◽  
Concentration Technique ◽  
Viverrini Infection

Opisthorchis viverrini infection is the major parasitic infection problem in Southeast Asian countries, and long-term infection will lead to cholangiocarcinoma (CCA), the bile duct cancer. The early diagnosis of O. viverrini infection may interrupt the progression of the opisthorchiasis and other related illnesses, especially CCA. The current diagnostic procedure is stool examination by microscope-based methods such as direct smear and concentration techniques but it is limited by low parasite egg numbers. The molecular diagnosis prompts the chance to evaluate the light infection with low number of parasite eggs but is currently inconvenient for routine use due to special equipment requirement and unstable sensitivities. Our present study aims to establish the efficiency of OvNad subunits, the mitochondrial gene, for introducing as a potential diagnostic target by conventional PCR, the cheapest and easiest molecular procedure. A total of 166 stool samples were investigated microscopically by the PBS-ethyl acetate concentration technique (PECT); 75 samples were O. viverrini positive with 28 samples that were positive with single parasite (hookworm, A. lumbricoides, S. stercoralis, Taenia spp., and T. trichiura), 11 samples were with mixed infection, and 52 samples were without parasite detection. The detection limits of OvNad subunits were evaluated in artificially spiked samples containing 0, 1, 5, 10, 20, 50, and 100 Ov-eggs. The result suggested that the best detection efficacy was of OvNad5 that had exact detection limits at only 5 eggs. In the PCR amplification of OvNad subunits, there exist 100% specificities with varied sensitivities from 64%, 88%, 80%, and 100% of OvNad1, OvNad2, OvNad4, and OvNad5, respectively. OvNad subunits were amplified specifically without cross reactivity with the other collected parasites. Our study established that OvNad subunits, especially OvNad5, are the potent candidates for PCR amplification of stool containing Ov-eggs with high confidential sensitivity, specificity, PPV, and NPV even in the light infection that would be a benefit for developing as a routine diagnosis of O. viverrini infection.

scholarly journals Hemocyte subpopulation changes in Bithynia snails infected with Opisthorchis viverrini in Thailand.

2019 ◽  
Author(s):  
Kulwadee Suwannatrai ◽  
Apiporn Thinkhamrop Suwannatrai ◽  
Chalermlap Donthaisong ◽  
Patpicha Arunsan ◽  
Kavin Thinkhamrop ◽  
...  
Keyword(s):  
Host Species ◽  
Parasitic Infection ◽  
Opisthorchis Viverrini ◽  
Parasitic Infections ◽  
Defense System ◽  
Intermediate Hosts ◽  
Molecular Functions ◽  
Viverrini Infection

The Bithynia snails of B. funiculata, B. siamensis siamensis and B. siamensis goniomphalos, are first intermediate hosts of Opisthorchis viverrini. The success of parasitic infection in snails is related to the host species and efficiency of their internal defense system. Parasitic infections in snails exhibited significant variations in the number of hemocytes in hemolymph. This study investigated hemocyte counts in Bithynia snails for various durations of O. viverrini infection. The three species/subspecies snails were infected with O. viverrini and hemocytes were counted at post-exposed periods of 2, 4, 6, 12, 24, 48, 96 h, and 1, 4 and 8 wk to evaluate the hemocyte response to the infection. Three major hemocyte morphotypes, namely, agranulocytes, semigranulocytes and granulocytes, were observed. After infection, the differential hemocyte morphotype counts in all species/subspecies significantly increased in granulocytes at 2 h (p < 0.05) and decreased in semigranulocytes and agranulocytes at 2 to 4 h (p < 0.05). The total hemocyte counts were significant increased at 2 h after exposure in B. s. siamensis and B. funiculata (p < 0.05). Additionally, the number of hemocytes was reduced after exposure at 4, 6 and 12 h in B. s. goniomphalos and at 12, 24 and 48 h in B. funiculata (p < 0.05). From 96 h to 8 wk, the number of hemocytes was reestablished in the hemolymph, indicating that defensive cells in the host species have different mechanisms for their susceptibility or resistance to parasitic infections. Further studies on molecular functions will be done.

scholarly journals Applicability of human-specific STR systems, GlobalFiler™ PCR Amplification Kit, Investigator 24plex QS Kit, and PowerPlex® Fusion 6C in chimpanzee (Pan troglodytes)

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Abhishek Singh ◽  
Vivek Sahajpal ◽  
Mukesh Thakur ◽  
Lalit Kumar Sharma ◽  
Kailash Chandra ◽  
...  
Keyword(s):  
Population Genetics ◽  
Pan Troglodytes ◽  
Human Population ◽  
Pcr Amplification ◽  
Forensic Analysis ◽  
Cross Reactivity ◽  
Allele Size ◽  
Male And Female ◽  
Successful Amplification ◽  
Human Specific

Abstract Objectives Human identification systems based on STRs are widely used in human population genetics and forensic analysis. This study aimed to validate the cross-reactivity of three widely known human-specific STR identification systems i.e. GlobalFiler™ PCR Amplification Kit, Investigator 24plex QS Kit, and PowerPlex® Fusion 6C in chimpanzee. Results The present study revealed the successful amplification of 18 loci using GlobalFiler™ PCR Amplification Kit, 18 loci using Investigator 24plex QS Kit, and 20 loci using PowerPlex® Fusion 6C system. The marker Amelogenin (AMEL) showed differential allele size between male and female revealing the gender identity of chimpanzees and thus validates their application concerning forensic examination, population estimation, and genetic analysis.

Reversal of biliary tract abnormalities associated with Opisthorchis viverrini infection following praziquantel treatment

1993 ◽  
Vol 87 (2) ◽  
pp. 194-197 ◽  
Author(s):  
Eimorn Mairiang ◽  
Melissa R. Haswell-Elkins ◽  
Pisaln Mairiang ◽  
Paiboon Sithithaworn ◽  
David B. Elkins
Keyword(s):  
Biliary Tract ◽  
Opisthorchis Viverrini ◽  
Praziquantel Treatment ◽  
Viverrini Infection

Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit beta as a potential biomarker for Opisthorchis viverrini infection and cholangiocarcinoma

Parasitology ◽  
2021 ◽  
pp. 1-35
Author(s):  
Sattrachai Prasopdee ◽  
Yodying Yingchutrakul ◽  
Sittiruk Roytrakul ◽  
Montinee Pholhelm ◽  
Teva Phanaksri ◽  
...  
Keyword(s):  
Opisthorchis Viverrini ◽  
Catalytic Subunit ◽  
Potential Biomarker ◽  
Viverrini Infection

scholarly journals Intestinal parasitic infections among HIV infected patients on antiretroviral therapy attending Debretabor General Hospital, Northern Ethiopia: a cross-sectional study

2020 ◽  
Author(s):  
Ermias Alemayehu ◽  
Alemu Gedefie ◽  
Aderaw Adamu ◽  
Jemal Mohammed ◽  
Brhanu Kassanew ◽  
...  
Keyword(s):  
General Hospital ◽  
Intestinal Parasites ◽  
Parasitic Infection ◽  
Cross Sectional Study ◽  
Parasitic Infections ◽  
Northern Ethiopia ◽  
Intestinal Parasitic Infection ◽  
Sectional Study ◽  
Cross Sectional ◽  
Concentration Technique

Abstract Background: Parasitic infections are known causes of morbidity among HIV infected patients with low CD4+ counts who are on antiretroviral therapy; mainly as a result of immune suppression. Thus, this study aimed to assess the extent of intestinal parasitic infection and its related risk factors among HIV infected patients attending ART clinic of Debretabor General Hospital, Northern Ethiopia.Patients and methods: A health facility based cross sectional study was conducted on a total of 383 HIV infected patients attending ART clinic of Debretabor General Hospital, Northern Ethiopia from December 2018 to March 2019. An interview based structured questionnaire were used to gather socio-demographic and risk factor data. About 5 grams of fresh stool specimen and 4 ml of venous blood sample were collected, then transported and tested in accordance with the laboratory standard operating procedures. The obtained data was entered into SPSS version 22.0; and analyzed. P-value <0.05 with 95% confidence interval was considered statistically significant.Result: The overall prevalence of intestinal parasites was 25.3%; with 18% and 23.8% by direct wet mount and formol-ether concentration technique respectively. 8 (2.1%) patients were infected by multiple parasites in concentration technique. Ascaris lumbricoides, 23(25.3%) was the most frequently identified parasite. Parasitic infection was significantly higher among illiterates (P=0.011); patients with a CD4 count of <200 cells/mm3 (P<0.001) and among patients who did not have latrine (P=0.049) than their counter parts.Conclusion: Relatively higher prevalence of intestinal parasitic infection was found among HIV/AIDS patients. The distribution of intestinal parasites is greatly affected by illiteracy, reduced CD4+ counts and absence of toilet. Thus, HIV/AIDS patients with low CD4+ counts should be diagnosed consistently for intestinal parasites and routine stool examination and awareness creation should be advocated to be included as an essential component of the ART monitoring strategy for improved patient care.

scholarly journals Selection and Identification of Novel Aptamers Specific for Clenbuterol Based on ssDNA Library Immobilized SELEX and Gold Nanoparticles Biosensor

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2337 ◽  
Author(s):  
Xixia Liu ◽  
Qi Lu ◽  
Sirui Chen ◽  
Fang Wang ◽  
Jianjun Hou ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Limit Of Detection ◽  
Retention Rate ◽  
Pcr Amplification ◽  
Cross Reactivity ◽  
Enriched Library ◽  
Amplification Curve ◽  
Specificity And Sensitivity

We describe a multiple combined strategy to discover novel aptamers specific for clenbuterol (CBL). An immobilized ssDNA library was used for the selection of specific aptamers using the systematic evolution of ligands by exponential enrichment (SELEX). Progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing. Candidate aptamers were picked and preliminarily identified using a gold nanoparticles (AuNPs) biosensor. Bioactive aptamers were characterized for affinity, circular dichroism (CD), specificity and sensitivity. The Q-PCR amplification curve increased and the retention rate was about 1% at the eighth round. Use of the AuNPs biosensor and CD analyses determined that six aptamers had binding activity. Affinity analysis showed that aptamer 47 had the highest affinity (Kd = 42.17 ± 8.98 nM) with no cross reactivity to CBL analogs. Indirect competitive enzyme linked aptamer assay (IC-ELAA) based on a 5′-biotin aptamer 47 indicated the limit of detection (LOD) was 0.18 ± 0.02 ng/L (n = 3), and it was used to detect pork samples with a mean recovery of 83.33–97.03%. This is the first report of a universal strategy including library fixation, Q-PCR monitoring, high-throughput sequencing, and AuNPs biosensor identification to select aptamers specific for small molecules.

scholarly journals A Rapid and International Applicable Diagnostic Device for Cobra (Genus Naja) Snakebites

Toxins ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 572
Author(s):  
Jing-Hua Lin ◽  
Wang-Chou Sung ◽  
Jiunn-Wang Liao ◽  
Dong-Zong Hung
Keyword(s):  
Detection Limits ◽  
Rapid Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunochromatographic Test ◽  
Tissue Destruction ◽  
Diagnostic Device ◽  
Life Threatening ◽  
The Cross ◽  
Geographical Characteristics

Cobra snakes (genus Naja) are some of the most dangerous snake species in Asia and Africa, as their bites cause severe life-threatening respiratory failure and local tissue destruction, especially in the case of late diagnosis. The differential diagnosis of snakebite envenomation still mainly relies upon symptomatology, the patient’s description, and the experience of physicians. We have designed a rapid test, immunochromatographic test of cobra (ICT-Cobra), which obtained fair results in improving the diagnosis and treatment of Naja (N.) atra snakebites in Taiwan. In this study, we further investigated the feasibility of applying the kit for the detection of other cobra venoms based on the potential interspecies similarity. We firstly demonstrated the cross-reactivity between eight venoms of medically important cobra species and the rabbit anti-N. atra IgG that was used in ICT-Cobra by Western blotting and sandwich enzyme-linked immunosorbent assay. Then, ICT-Cobra was used to detect various concentrations of the eight venoms to elucidate its performance. Noticeable correlations between the cross-reactivity of venoms from genus Naja snakes and existing geographical characteristics were found. ICT-Cobra could detect venoms from other Asian cobras with variable detection limits comparable to those observed for N. atra, but the kit was less successful in the detection of venom from African cobras. The similar but slightly different venom components and the interaction between venom and rabbit anti-N. atra IgG led to variations in the detection limits. The transcontinental usage of ICT-Cobra might be possible due to the cross-reactivity of antibodies and similarities among the larger-sized proteins. This study showed that the close immunological relationships in the genus Naja could be used to develop a venom detection kit for the diagnosis of cobra envenomation in both Asian and African regions. Additional clinical studies and technical adjustments are still needed to improve the efficacy and broadening the application of ICT-Cobra in the future.

scholarly journals Uncooked fish consumption among those at risk of Opisthorchis viverrini infection in central Thailand

PLoS ONE ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. e0211540 ◽  
Author(s):  
Picha Suwannahitatorn ◽  
Joanne Webster ◽  
Steven Riley ◽  
Mathirut Mungthin ◽  
Christl A. Donnelly
Keyword(s):  
At Risk ◽  
Fish Consumption ◽  
Opisthorchis Viverrini ◽  
Central Thailand ◽  
Viverrini Infection

Asymptomatic giardiasis-more prevalent in refugees than in native inhabitants of the city of Nis, Serbia

Open Medicine ◽  
2008 ◽  
Vol 3 (2) ◽  
pp. 203-206
Author(s):  
Natasa Miladinovic-Tasic ◽  
Suzana Tasic ◽  
Ivana Kranjcic-Zec ◽  
Gordana Tasic ◽  
Aleksandar Tasic ◽  
...  
Keyword(s):  
Microscopic Examination ◽  
Gold Standard ◽  
Parasitic Infection ◽  
Giardia Duodenalis ◽  
Population Group ◽  
Concentration Technique ◽  
Large Numbers ◽  
Stool Samples ◽  
Statistical Survey ◽  
The City

AbstractGiardiasis is a parasitic infection of the digestive tract, most commonly occurring in closed communities such as schools, kindergartens, prisons, and campuses. The civil war in the former Yugoslav republics and in Kosovo caused a large number of refugees to take shelter in the territory of Serbia. Such large numbers of refugees could be accommodated only in the collective centers. Our aim was to examine the differences in the prevalence of asymptomatic giardiasis among 122 refugees from the former Yugoslav republics who lived in the collective centers in Nis, Serbia, and 241 native Nis inhabitants. Conventional microscopic examination (CME) of three stool samples with or without concentration technique and the enzyme immunoassay (EIA) methods were used. The CME method of three stool samples is considered the gold standard in our statistical survey. Asymptomatic giardiasis is found in 7 refugees (5.7%) using the EIA method, while using the CME (3 samples) Giardia duodenalis (G. duodenalis) was detected in 6 persons (4.9%). Using the EIA method and the CME (3 samples) G. duodenalis was detected in only 1 person in the population group of native inhabitants (0.4%). Asymptomatic giardiasis was more prevalent in the population group of refugees accommodated in collective centers than in native inhabitants in the Nis municipality, Serbia.

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