scholarly journals MOLECULAR DIAGNOSIS AND PHYLOGENETIC ANALYSIS OF BABESIA SPECIES ISOLATED FROM TICKS OF INFESTED CATTLE IN WASIT GOVERNORATE, IRAQ

2021 ◽  
Vol 52 (1) ◽  
pp. 136-145
Author(s):  
Al-Abedi & Al-Amery

The aim of current study is to detect Babesia bovis, B. bigemina, and B. divergens in ticks using molecular polymerase chain reaction (PCR) assay. In a totally 180 cattle examined to collect of tick samples during December 2018 to August 2019, the findings were revealed on 63 (35%) cattle infested with ticks that classified morphologically to belong to the genus of Hyalomma and genus of Rhipicephalus. From 50 tick samples tested by PCR assay, 41 (82%) were infested by Babesia genus including 30 (68.18%) infested with B. bovis and 11 (31.82%) infested with B. bigemina; whereas, no tick samples were found to be infested with B. divergens. To document the local isolated strains, five PCR products of each B. bovis and B. bigemina positive strains were selected, sequenced and reported in the NCBI under the accession numbers of (MN727083.1, MN727084.1, MN727085.1, MN727086.1, and MN727087.1) and (MN741113.1, MN741114.1, MN741115.1,MN741116.1, and MN741117.1) respectively.

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.


2020 ◽  
Vol 17 (1) ◽  
Author(s):  
Yang Zhang ◽  
Chunyang Dai ◽  
Huiyan Wang ◽  
Yong Gao ◽  
Tuantuan Li ◽  
...  

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.


Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.


1994 ◽  
Vol 66 (1) ◽  
pp. 69-75 ◽  
Author(s):  
Toshimichi Yamamoto ◽  
Keiji Tamaki ◽  
Toshinori Kojima ◽  
Rieko Uchihi ◽  
Yoshinao Katsumata ◽  
...  

2016 ◽  
Vol 4 (2) ◽  
pp. 264-270 ◽  
Author(s):  
Aml Soliman ◽  
Asmaa Abdel Aal ◽  
Reham Afify ◽  
Noha Ibrahim

AIM: Aim was to detect Brain and Acute Leukemia, Cytoplasmic (BAALC) and ETS-related gene (ERG) expression in patients with acute myeloid leukemia (AML) as well as to study their biologic and prognostic impact on the disease outcome and survival.PATIENTS AND METHODS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression.RESULTS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression. BAALC was expressed in 36 (81.82%) of AML cases versus 10 (22.72%) of the control group which was highly statistically significant (P < 0.001). While ERG was positive in 39(88.64%) of cases and 8(18.18 %) of controls and that was also highly statistically significant (P < 0.001).CONCLUSION: Further researches still needed to clarify the role of BAALC and ERG in the pathogenesis of leukemia and their importance as targets for treatment of AML.


1996 ◽  
Vol 8 (2) ◽  
pp. 181-185 ◽  
Author(s):  
Patricia K. Holyoake ◽  
Gary F. Jones ◽  
Peter R. Davies ◽  
Dennis L. Foss ◽  
Michael P. Murtaugh

A polymerase chain reaction (PCR) assay was used to confirm the presence of ileal symbiont (IS) intracellularis in 3 swine herds with a history of proliferative enteritis (PE). Two pooled fecal specimens, each comprising 5 individual stool samples, were collected from pen floors to screen for the presence of IS intracellularis and determine the age range of pigs shedding the organism. IS intracellularis was detected in the feces of clinically normal 10–25week-old grower/finisher pigs, indicating that this age range of pigs was the main source of infection for younger nursery pigs. Shedding continued without clinical disease when 10–100 g/ton of tylosin or 10 g/ton of chlortetracycline was added to the feed. PCR testing of pooled fecal samples can be used to identify groups of pigs affected with PE. The results of this study indicate that this PCR assay has the potential to accurately assess the IS intracellularis infection status of swine herds and the association of IS intracellular-is with PE and growth performance.


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