scholarly journals Molecular Confirmation of Salmonella typhimuriumin Poultry from Kathmandu Valley

2018 ◽  
Vol 4 ◽  
pp. 86-89 ◽  
Author(s):  
Sanjeev Kumar Adhikari ◽  
Arrogya Gyawali ◽  
Sajan Shrestha ◽  
Swoyam Prakash Shrestha ◽  
Meera Prajapati ◽  
...  

A prevalence study was carried to isolate Salmonella typhimurium from blood (n= 50) and gut samples (n=100) of poultry in Kathmandu valley during early 2016. Salmonella typhimurium bacteria isolated in the selective media were biochemically confirmed based on Bergey’s Manual. Two sets of oligonucleotide primers-the genus specific 16S rRNA and the organism specific invA were employed for molecular level confirmation by the Polymerase Chain Reaction (PCR) assay. The amplified fragments in 1% agarose gel observed at 406bp and 285bp, respectively confirmed the isolates to be Salmonella typhimurium. Of 150 samples tested, Salmonella typhimurium were isolated from 49 samples, among which nine were from blood (18%) and forty from the gut (40%). The present result indicated an alarmingly high level of Salmonella typhimurium, which can result inzoonotic infection in humans owing to increased contact with poultry and consumption of poultry products in the Kathmandu valley.

1999 ◽  
Vol 10 (10) ◽  
pp. 646-651 ◽  
Author(s):  
T Beattie ◽  
A Moyes ◽  
C Patrizio ◽  
H Young

Tetracycline resistant Neisseria gonorrhoeae (TRNG) contain a 25.2 MDa TetM plasmid encoding a 68KDa cytoplasmic protein which confers high-level tetracycline resistance. The aim of this study was to subtype all TRNG isolated in Scotland between 1992 and 1998. Subtyping was performed by a polymerase chain reaction (PCR) assay which characterizes the TetM plasmid as either the Dutch variant (443 base pair product) or the American variant (777 base pair product). Of the 78 TRNG isolates, 35 were the American variant and 43 were the Dutch variant. TRNG were distributed amongst 30 serovar/auxotype classes, the most common being 1A6/NR (11.5%), 1A6/P (14.1%) and 1B4/NR (14.1%). The country where infection was acquired was known for 36 of the 46 TRNG strains isolated between 1996 and 1998. All infections acquired in Asia and South America were the Dutch variant whereas all infections acquired in Africa were the American variant. A penicillinase plasmid was present in 66% (23/35) of the American variant TRNG compared with 51% (22/43) of the Dutch variant: the 3.2 MDa penicillinase plasmid was found in 87% of the American variant TRNG whereas the 4.4 MDa penicillinase plasmid was found in 68% of the Dutch variant TRNG. We conclude that subtyping of TRNG by PCR is a useful tool in studying the epidemiology of gonococcal infection due to plasmid-mediated resistant isolates.


2010 ◽  
Vol 9 (3) ◽  
pp. 97
Author(s):  
A. A. K. Jawad, A. H. Al-Hamadani And H. A. M. Al-Karawy

A total of 480 fecal samples were collected from children (less than 3 years old) , ofboth sexes suffering from diarrhea who admitted to The Teaching Hospital of Maternity andPediatrics in Al- Diwaniya governorate, Salmonella spp. were isolated and identified usingbacterial culturing on selective media, in addition to, biochemical and Mini API 20E andserotyping by monovalent antisera. Polymerase chain reaction (PCR) was used to detect fimCgene encoding for biosynthesis of fimC of Salmonella typhimurium. The results revealed thatthe rate of Salmonella isolates in fecal samples of patients were (38/480) 7.9% using culturaland Mini API20 E, The results of serotyping revealed that isolates there were 34 belong toSalmonella spp. Of these isolates 30 belong to S . typhimurium, while the remainingbelong to S. enteritidis ( 2 isolates) and S. meunchen (2 isolates), when the PCR techniquewas used to detect the presence of fimC gene, 32 Salmonella isolates were belong to S.typhimurium appeared to contain this gene . The results of this study revealed that thePCR technique had a high specifity (100%) in detection of S. typhimurium in comparison toserotyping.


2020 ◽  
Vol 8 (2) ◽  
pp. 716
Author(s):  
Wanessa Shuelen Costa Araújo ◽  
Carina Martins Moraes ◽  
Vanderson Vasconcelos Dantas ◽  
Andrey Carlos Sacramento de Oliveira ◽  
Talita Bandeira Roos ◽  
...  

Pirarucu is a freshwater fish that presents a great commercial value for being well accepted by the consumers and for showing excellent meat quality. The zoological identification of fisheries during industrial processing is harmed by the removal of external morphologic characteristics, facilitating fraudulent practices in commercialization. In this context, the identification at the molecular level is an important tool in the inspection and commercialization of the fishery. The DNA resists to the processing methods, like the salting, the most common way of commercializing the pirarucu. The Polymerase chain reaction, PCR assay, were applied in samples that suffered degradation or have gone under industrialization methods. This work aimed use the PCR technique as a tool to authenticate the Arapaima gigas species and to avoid possible frauds in commercially available products. The obtained data showed the efficiencies of the DNA extraction, the amplification of the target sequence, and identification of the genetic material through PCR. It is possible to conclude that the PCR technique that was standardized in the present study showed high sensibility, precision, and specificity for the detection of the genetic material of Arapaima gigas, constituting a useful tool for the monitoring and inspection during its commercialization.


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.


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.


Author(s):  
Deborah Marriott ◽  
Rohan Beresford ◽  
Feras Mirdad ◽  
Damien Stark ◽  
Allan Glanville ◽  
...  

Abstract Our Australian hospital tested almost 22 000 symptomatic people over 11 weeks for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in a multiplex polymerase chain reaction (PCR) assay. Following travel bans and physical distancing, SARS-CoV-2 and other respiratory viruses diagnoses fell dramatically. Increasing rhinovirus diagnoses as social control measures were relaxed may indirectly indicate an elevated risk of coronavirus disease 2019 (COVID-19) resurgence.


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