Epidemics of schistosomiasis in military staff assigned to endemic areas: standard diagnostic techniques and the development of real-time PCR techniques

The present study was carried out to investigate the involvement of different bovine papillomaviruses in the teat warts of cattle. A total of 11 teat wart samples showing rice grain-like and small, sessile elevated greyish or flesh-like growths were collected from dairy cattle. DNA was extracted from these teat wart samples and PCR and real time PCR techniques were applied using specific primers for BPV-1 and -10 to detect the presence of viral nucleic acid. PCR revealed the presence of viral DNA of BPV-1 and -10 in three and seven samples, respectively. Quantification using real time PCR revealed that the copy numbers of the viral DNA of BPV-1 and -10 DNA varied from 1.12E + 04 to 2.99E + 04 and 3.56E + 02 to 5.23E + 06, respectively. From the present study it can be concluded that BPV-1 and -10 are involved in production of rice grain-like and sessile elevated growths on the teats of cattle.

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Analysis of the structure of microbial community in soils with different degrees of salinization using T-RFLP and real-time PCR techniques

Eurasian Soil Science ◽

10.1134/s1064229312020044 ◽

2012 ◽

Vol 45 (2) ◽

pp. 147-156 ◽

Cited By ~ 35

Author(s):

E. E. Andronov ◽

S. N. Petrova ◽

A. G. Pinaev ◽

E. V. Pershina ◽

S. Zh. Rakhimgalieva ◽

...

Keyword(s):

Microbial Community ◽

Real Time ◽

Real Time Pcr ◽

Pcr Techniques

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Field Validation of SYBR Green- and TaqMan-Based Real-Time PCR Using Biopsy and Swab Samples To Diagnose American Tegumentary Leishmaniasis in an Area Where Leishmania (Viannia) braziliensis Is Endemic

Journal of Clinical Microbiology ◽

10.1128/jcm.01954-16 ◽

2016 ◽

Vol 55 (2) ◽

pp. 526-534 ◽

Cited By ~ 22

Author(s):

Ciro Martins Gomes ◽

Mariana Vicente Cesetti ◽

Natália Aparecida de Paula ◽

Sebastián Vernal ◽

Gaurav Gupta ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Sybr Green ◽

Conventional Pcr ◽

Content Type ◽

Tegumentary Leishmaniasis ◽

Precise Diagnosis ◽

American Tegumentary Leishmaniasis ◽

Pcr Techniques ◽

Composite Reference Standard

ABSTRACTThe precise diagnosis of American tegumentary leishmaniasis (ATL) is an essential task due to the disease's associated morbidity. A noninvasive, extremely sensitive, and highly specific exam is critical, particularly for mucosal leishmaniasis (ML), in which a low parasite quantity is expected. We aimed to compare the diagnostic accuracy of swab and biopsy sample analysis using SYBR Green- and TaqMan-based real-time PCR (qPCR) assays with that of a composite reference standard consisting of the Montenegro skin test, serology, histopathology, smears, culture, and conventional PCR. In total, 55 patients with ATL (ML, 18 patients; cutaneous leishmaniasis [CL], 37 patients) and 36 patients without ATL were studied. qPCR analysis of swabs was more accurate when using SYBR Green (87.88%; 95% confidence interval [CI], 77.86 to 93.73 patients) than when using TaqMan (78.79%; 95% CI, 67.49 to 86.92%) (P= 0.031). SYBR Green (84.72%; 95% CI, 74.68 to 91.25%) was also more accurate than TaqMan (73.61%; 95% CI, 62.42 to 82.41%) for biopsy samples (P= 0.008). All qPCR methods were 100% specific. Swabs and biopsy specimens had similar sensitivity when using the same chemistry (P= 0.125 for SYBR Green andP= 0.625 for TaqMan). Moreover, qPCR achieved better performance than most existing techniques used for the diagnosis of ATL and also detected theLeishmaniaparasite in a greater proportion of patients than the associated histopathology, smear, culture, and conventional PCR techniques did. Swabs therefore represent a useful diagnostic tool because they not only are noninvasive but also can achieve an accuracy similar to that of biopsy samples. The high accuracy of SYBR Green-based qPCR may also reduce the requirement for associated parasitological tests for ATL diagnosis.

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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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The Mbam drainage system and onchocerciasis transmission post ivermectin mass drug administration (MDA) campaign, Cameroon

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0008926 ◽

2021 ◽

Vol 15 (1) ◽

pp. e0008926

Author(s):

Raphael Awah Abong ◽

Glory Ngongeh Amambo ◽

Ali Ahamat Hamid ◽

Belinda Agbor Enow ◽

Amuam Andrew Beng ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Large Scale ◽

Water Discharge ◽

Drainage System ◽

River System ◽

High Water ◽

Diagnostic Techniques ◽

Dry Seasons ◽

The Impact

Background The impact of large scale Mass Drug Adminstration (MDA) of ivermectin on active onchocerciasis transmission by Simulium damnosum, which transmits the parasite O. volvulus is of great importance for onchocerciasis control programmes. We investigated in the Mbam river system area, the impact of MDA of ivermectin on entomological indices and also verify if there are river system factors that could have favoured the transmission of onchocerciasis in this area and contribute to the persistence of disease. We compared three independent techniques to detect Onchocerca larvae in blackflies and also analyzed the river system within 9 months post-MDA of ivermectin. Method Simulium flies were captured before and after 1, 3, 6 and 9months of ivermectin-MDA. The biting rate was determined and 41% of the flies dissected while the rest were grouped into pools of 100 flies for DNA extraction. The extracted DNA was then subjected to O-150 LAMP and real-time PCR for the detection of infection by Onchocerca species using pool screening. The river system was analysed and the water discharge compared between rainy and dry seasons. Principal findings We used human landing collection method (previously called human bait) to collect 22,274 adult female Simulium flies from Mbam River System. Of this number, 9,134 were dissected while 129 pools constituted for molecular screening. Overall biting and parous rates of 1113 flies/man/day and 24.7%, respectively, were observed. All diagnostic techniques detected similar rates of O. volvulus infection (P = 0.9252) and infectivity (P = 0.4825) at all monitoring time points. Onchocerca ochengi larvae were only detected in 2 of the 129 pools. Analysis of the river drainage revealed two hydroelectric dams constructed on the tributaries of the Mbam river were the key contributing factor to the high-water discharge during both rainy and dry seasons. Conclusion Results from fly dissection (Microscopy), real-time PCR and LAMP revealed the same trends pre- and post-MDA. The infection rate with animal Onchocerca sp was exceptionally low. The dense river system generate important breeding sites that govern the abundance of Simulium during both dry and rainy seasons.

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