Multiplex real-time PCR revealed very high prevalence of soil-transmitted helminth infections among aborigines in Peninsular Malaysia

Differently from alpha- and betaherpesviruses affecting swine, interest in the recently discovered Suid gammaherpesvirus 3, Suid gammaherpesvirus 4, and Suid gammaherpesvirus 5, also known as porcine lymphotropic herpesviruses (PLHV-1, PLHV-2, and PLHV-3), has largely focused on their role as potential zoonotic agents in cases of xenotransplantation. However, their role as primary pathogens of swine or as co-factors for other lymphotropic infections has essentially been neglected. The present study aims at filling this gap, evaluating the association between PLHVs infection and different clinical conditions and/or porcine circovirus (PCV) co-infection. One hundred seventy-six samples were obtained from different animals located in a high-density pig area of Northern Italy in the period 2017–2020. The presence of PLHVs and PCVs was tested and quantified by specific real-time PCR: PLHVs were widespread among pigs (PLHV-1, PLHV-2, and PLHV-3 prevalence was 28.97%, 10.79%, and 4.54%, respectively) and detected in all considered tissues and clinical conditions. Frequent co-infections were also observed among PLHVs and with PCVs, although a significant association was not detected with the exception of a positive interaction between PLHV-1 and PLHV-3, and a negative one between PLHV-2 and PCV-2. Significantly, no association between PLHVs, alone or in co-infection, emerged with any of the considered clinical signs, their frequency being comparable between healthy and diseased animals. Based on these pieces of evidence and despite their high prevalence, PLHVs’ relevance for the swine industry appears negligible, either as primary pathogens or as predisposing factors for circovirus-induced diseases.

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Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

10.21203/rs.3.rs-18430/v1 ◽

2020 ◽

Author(s):

Vu Thuy Duong ◽

Le Thi Phuong Tu ◽

Ha Thanh Tuyen ◽

Le Thi Quynh Nhi ◽

James I Campbell ◽

...

Keyword(s):

Escherichia Coli ◽

Real Time ◽

Real Time Pcr ◽

Clinical Samples ◽

Middle Income ◽

Middle Income Countries ◽

Virulence Markers ◽

Low Middle Income Countries ◽

High Prevalence ◽

Pcr Assays

Abstract BackgroundDiarrhoeagenic Escherichia coli (DEC) infections are common in children in low-middle income countries (LMICs). However, detecting the various DEC pathotypes is complex as they cannot be differentiated by classical microbiology. We developed four multiplex real-time PCR assays were to detect virulence markers of six DEC pathotypes; specificity was tested using DEC controls and other enteric pathogens. PCR amplicons from the six E. coli pathotypes were purified and amplified to be used to optimize PCR reactions and to calculate reproducibility. After validation, these assays were applied to clinical samples from healthy and diarrhoeal Vietnamese children and associated with clinical data. ResultsThe multiplex real-time PCRs were found to be reproducible, and specific. At least one DEC variant was detected in 34.7% (978/2,815) of the faecal samples from diarrhoeal children; EAEC, EIEC and atypical EPEC were most frequent Notably, 41.2% (205/498) of samples from non-diarrhoeal children was positive with a DEC pathotype. In this population, only EIEC, which was detected in 34.3% (99/289) of diarrhoeal samples vs. 0.8% (4/498) non-diarrhoeal samples (p<0.001), was significantly associated with diarrhoea. Multiplex real-time PCR when applied to clinical samples is an efficient and high-throughput approach to DEC pathotypes. ConclusionsThis approach revealed high carriage rates of DEC pathotypes among Vietnamese children. We describe a novel diagnostic approach for DEC, which provides baseline data for future surveillance studies assessing DEC burden in LMICs.

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High prevalence of intestinal parasite infestations among stunted and control children aged 2 to 5 years old in two neighborhoods of Antananarivo, Madagascar

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0009333 ◽

2021 ◽

Vol 15 (4) ◽

pp. e0009333

Author(s):

Azimdine Habib ◽

Lova Andrianonimiadana ◽

Maheninasy Rakotondrainipiana ◽

Prisca Andriantsalama ◽

Ravaka Randriamparany ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Large Scale ◽

Intestinal Parasite ◽

Detection Rates ◽

Quality Of Water ◽

High Prevalence ◽

Antiparasitic Drugs ◽

Associated Risk Factors ◽

And Control

Background This study aimed to compare the prevalence of intestinal parasite infestations (IPIs) in stunted children, compared to control children, in Ankasina and Andranomanalina Isotry (two disadvantaged neighborhoods of Antananarivo, Madagascar), to characterize associated risk factors and to compare IPI detection by real-time PCR and standard microscopy techniques. Methodology/Principal findings Fecal samples were collected from a total of 410 children (171 stunted and 239 control) aged 2–5 years. A single stool sample per subject was examined by simple merthiolate-iodine-formaldehyde (MIF), Kato-Katz smear and real-time PCR techniques. A total of 96.3% of the children were infested with at least one intestinal parasite. The most prevalent parasites were Giardia intestinalis (79.5%), Ascaris lumbricoides (68.3%) and Trichuris trichiura (68.0%). For all parasites studied, real-time PCR showed higher detection rates compared to microscopy (G. intestinalis [77.6% (n = 318) versus 20.9% (n = 86)], Entamoeba histolytica [15.8% (n = 65) versus 1.9% (n = 8)] and A. lumbricoides [64.1% (n = 263) versus 50.7% (n = 208)]). Among the different variables assessed in the study, age of 4 to 5 years (AOR = 4.61; 95% CI, (1.35–15.77)) and primary and secondary educational level of the mother (AOR = 12.59; 95% CI, (2.76–57.47); AOR = 9.17; 95% CI, (2.12–39.71), respectively) were significantly associated with IPIs. Children drinking untreated water was associated with infestation with G. intestinalis (AOR = 1.85; 95% CI, (1.1–3.09)) and E. histolytica (AOR = 1.9; 95% CI, (1.07–3.38)). E. histolytica was also associated with moderately stunted children (AOR = 0.37; 95% CI, 0.2–0.71). Similarly, children aged between 4 and 5 years (AOR = 3.2; 95% CI (2.04–5.01)) and living on noncemented soil types (AOR = 1.85; 95% CI, (1.18–2.09)) were associated with T. trichiura infestation. Conclusions/Significance The prevalence of IPIs is substantial in the studied areas in both stunted and control children, despite the large-scale drug administration of antiparasitic drugs in the country. This high prevalence of IPIs warrants further investigation. Improved health education, environmental sanitation and quality of water sources should be provided.

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Early determination of fetal sex in goat a comparison between real time PCR and ultrasonography

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/951/1/012059 ◽

2022 ◽

Vol 951 (1) ◽

pp. 012059

Author(s):

S A Hussein ◽

K M Karam

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Maternal Blood ◽

Fetal Sex ◽

Blood Specimens ◽

High Level ◽

Pcr Techniques ◽

Very High ◽

Genital Tubercle

Abstract The point of the current study is to assess the productivity of the real time PCR and ultrasound techniques in early determination of fetal sex in Iraqi singleton pregnant goats. Our investigation has been led in Iraq, Al-Diwanya city from 10/8/2020 – 15/1/2021. The examination incorporates 45 singleton pregnant Iraqi goats, which initially inspected by ultrasound to affirm pregnancy and to decide the fetal sex depending on the restriction of the genital tubercle of the goat fetuses, after that, blood specimens had been gathered from the jugular vein of all examined does to detect fetal sex by discovery of AMLX and SRY genes in the circling cells free fetal DNA (ccffDNA) in these maternal blood specimens by utilizing real time PCR. Our outcomes showed an exceptionally high level of accuracy in real time PCR in contrast with the ultrasound strategy. The outcomes were affirmed by the true fetal sex after parturition in the inspected does. The complete symptomatic rate were 51.11% (23/45) and 97.78% (44/45) for ultrasound and PCR strategies separately. The exactness level of genuine analyzed female and male caprine kidding were 58.33% (7/12), 48.48% (16/33), and 100% (12/12), 96.97% (32/33) for ultrasound and real time PCR techniques separately. While the exactness rates of the two techniques utilized in this investigation for early caprine fetal sexing in respect to early pregnancies periods analyzed uncovered 100% (13/13), 96.3% (26/27), 100% (5/5), and 61.54% (8/13), 40.74% (11/27), 80% (4/5) in early pregnancy periods (58-62, 63-67, 68-73) days for real time PCR and ultrasound strategies individually. In conclusion our outcomes revealed a huge predominant exactness and productivity in fetal sexing in Iraqi singleton pregnant does in early development periods, with very high accuracy in real time PCR in compare to ultrasound techniques.

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Real-time PCR Demonstrates High Prevalence of Schistosoma japonicum in the Philippines: Implications for Surveillance and Control

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0003483 ◽

2015 ◽

Vol 9 (1) ◽

pp. e0003483 ◽

Cited By ~ 28

Author(s):

Catherine. A. Gordon ◽

Luz P. Acosta ◽

Geoffrey N. Gobert ◽

Remigio M. Olveda ◽

Allen G. Ross ◽

...

Keyword(s):

Real Time ◽

Schistosoma Japonicum ◽

Real Time Pcr ◽

The Philippines ◽

High Prevalence ◽

And Control

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Performance of the commercial Schistosoma-Urine-CCA-test in comparison to Kato-Katz technique and Real-time PCR in a high prevalence region of Schistosoma mansoni at Lake Victoria, Tanzania

10.26226/morressier.572768e5d462b8029237f48c ◽

2016 ◽

Author(s):

Antje Fuss

Keyword(s):

Schistosoma Mansoni ◽

Real Time ◽

Real Time Pcr ◽

Lake Victoria ◽

High Prevalence

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Comparison of multiplex real-time PCR and ergosterol assays in quantifying Heterobasidion annosum in planta

Plant Protection Science ◽

10.17221/10507-pps ◽

2017 ◽

Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽

pp. 406-407

Author(s):

A.M. Hietala ◽

M. Eikenes ◽

M. Kvaalen ◽

H. Solheim ◽

C.G. Fossdal

Keyword(s):

Real Time ◽

Norway Spruce ◽

Real Time Pcr ◽

Comparative Method ◽

Heterobasidion Annosum ◽

Fungal Colonization ◽

In Planta ◽

Advantages And Disadvantages ◽

Very High

A quantitative multiplex real-time PCR procedure was developed to monitor the dynamics in Norway spruce-Heterobasidion annosum pathosystem. The assay reliably detected down to 1 pg of H. annosum DNA and 1 ng of host DNA in multiplex conditions. As a comparative method for quantifying fungal colonization, we applied the ergosterol assay. There was a very high correlation between the results obtained with the two methods, this strengthening the credibility of both assays. The advantages and disadvantages of these assays are discussed.

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