Prevalence of Salmonella typhi among Patients with Febrile Illness in Rural and Peri-Urban Populations of Vellore District, as Determined by Nested PCR Targeting the Flagellin Gene

2010 ◽  
Vol 14 (2) ◽  
pp. 107-112 ◽  
Author(s):  
Balaji Nandagopal ◽  
Sathish Sankar ◽  
Karthikeyan Lingesan ◽  
Kumarasekharan Chandrasekharan Appu ◽  
Baby Padmini ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Febrile Illness ◽  
Salmonella Typhi ◽  
Flagellin Gene ◽  
Urban Populations ◽  
Pcr Targeting

scholarly journals A Rapid and Sensitive Diagnosis of Typhoid Fever Based On Nested PCR-Voltammetric DNA Biosensor Using Flagellin Gene Fragment

2018 ◽  
Vol 16 (1) ◽  
pp. 87 ◽  
Author(s):  
Yeni Wahyuni Hartati ◽  
Santhy Wyantuti ◽  
M. Lutfi Firdaus ◽  
Nurul Auliany ◽  
Rini Surbakti ◽  
...  
Keyword(s):  
Typhoid Fever ◽  
Dna Sequences ◽  
Nested Pcr ◽  
Differential Pulse ◽  
Salmonella Typhi ◽  
Dna Biosensor ◽  
Pencil Graphite Electrode ◽  
Flagellin Gene ◽  
Complementary Sequence ◽  
Pcr Product

Typhoid fever caused by Salmonella typhi is an important issue for public health in the world. Laboratory methods for rapid and sensitive diagnosis are very important for disease management. The purpose of this study was to determine the performance of nested PCR–voltammetric DNA biosensor using flagellin gene (fla) of S. typhi as a marker. The differential pulse voltammetry using pencil graphite electrode was applied to measure the guanine oxidation signal of probes vs synthetic target stDNA and probes vs fla PCR product hybridizations. The probe DNA selectivity was examined by hybridized probes vs non-complementary sequence. The result showed that the first round nested PCR product can not be visualized by agarose electrophoresis, whereas using the voltammetric biosensor methods can be detected both for the first or second round nested PCR product. The average peak current of hybridized probe vs first and second round of PCR product was 2.32 and 1.47 μA respectively, at 0.9 V. Detection of the DNA sequences of the infectious diseases from PCR amplified real sample was also carried out using this voltammetric DNA biosensor methods.

scholarly journals Enhanced detection rate of typhoid fever among clinically suspected patients in a tertiary referral hospital in Dhaka, Bangladesh using nested polymerase chain reaction technology

2016 ◽  
Vol 41 (3) ◽  
pp. 138-143
Author(s):  
Shafinaz Khan ◽  
Ruhul Amin Mia ◽  
Sumaiya Khatun
Keyword(s):  
Polymerase Chain Reaction ◽  
Blood Culture ◽  
Typhoid Fever ◽  
Nested Pcr ◽  
Salmonella Typhi ◽  
Chain Reaction ◽  
Tertiary Referral Hospital ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Pcr Targeting

A nested polymerase chain reaction (PCR) specific for Salmonella enterica subspecies enteric serovar Typhi was used for the detection of the pathogen in blood. This study was done during the period of March 2013 to February 2014. A total of 80 clinically suspected cases of typhoid fever were included in the study. Blood was collected from all participating individuals. Nested PCR targeting the flagellin gene (fliC) of Salmonella Typhi & blood culture were done for each of the cases. The positivity rate of PCR & blood culture was 70%& 20% respectively. The positivity rate of PCR was significantly higher than blood culture (P< 0.05). With the nested PCR, S. Typhi DNAs were detected from blood specimens of 67.2% (43/64) patients among the suspected typhoid fever cases on the basis of clinical features but with negative cultures. We conclude that the PCR technique could be used as a novel diagnostic method of typhoid fever, particularly in culture-negative cases in an endemic country like Bangladesh.

scholarly journals Use of Urine with Nested PCR Targeting the Flagellin Gene (fliC) for Diagnosis of Typhoid Fever

2012 ◽  
Vol 50 (6) ◽  
pp. 1964-1967 ◽  
Author(s):  
Gopal Kumar ◽  
Chandra Bhan Pratap ◽  
Om Prakash Mishra ◽  
Kailash Kumar ◽  
Gopal Nath
Keyword(s):  
Typhoid Fever ◽  
Nested Pcr ◽  
Flagellin Gene ◽  
Pcr Targeting

scholarly journals Detection and partial genetic characterisation of novel avi- and siadenoviruses in racing and fancy pigeons (Columba livia domestica)

2016 ◽  
Vol 64 (4) ◽  
pp. 514-528 ◽  
Author(s):  
Mónika Z. Ballmann ◽  
Balázs Harrach
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Dna Polymerase ◽  
Nested Pcr ◽  
Columba Livia ◽  
Hexon Gene ◽  
Polymerase Gene ◽  
Viral Dna ◽  
Phylogeny Analysis ◽  
Pcr Targeting

Up to now, only a single adenovirus (AdV) isolate seemingly specific for pigeons, hence named pigeon AdV-1 (PiAdV-1), has been characterised at DNA sequence level. In the present work, the prevalence and diversity of AdVs occurring in domestic pigeon were examined by a survey performed on randomly collected samples using a very efficient, consensus nested PCR targeting the viral DNA polymerase gene. The newly detected viruses were characterised by sequencing and phylogeny analysis. Amplification of additional genome fragments was attempted by the use of several other PCR methods aiming at the hexon gene. During a 4-year survey, samples from dead or live, healthy pigeons originating from 27 lofts were examined in Hungary. Almost 50% of the samples (48 out of 97) proved to be positive for AdV. Sequence analysis revealed the presence of four hitherto unknown pigeon AdV types. PiAdV-1 was also identified in one sample. Two novel viruses named PiAdV-2 and -3 were found to belong to the genus Aviadenovirus, and two other novel types (PiAdV-4 and -5) to the genus Siadenovirus. This is the first report on the occurrence of siadenoviruses in birds belonging to the order Columbiformes. Approximately two-thirds of the PiAdV-2 genome was sequenced and analysed.

Nested PCR targeting intergenic spacer (IGS) in genotyping of Giardia duodenalis isolated from symptomatic and asymptomatic infected Egyptian school children

2016 ◽  
Vol 116 (2) ◽  
pp. 763-771 ◽  
Author(s):  
Eman M. Hussein ◽  
Ola A. Ismail ◽  
Amira B. Mokhtar ◽  
Samer E. Mohamed ◽  
Rania M. Saad
Keyword(s):  
School Children ◽  
Nested Pcr ◽  
Giardia Duodenalis ◽  
Intergenic Spacer ◽  
Pcr Targeting

scholarly journals Presentation and Treatment Outcomes of Liberian Children Age 5 Years and Under Diagnosed With Severe Malaria

2019 ◽  
Vol 6 ◽  
pp. 2333794X1988481
Author(s):  
Benetta Collins-Andrews ◽  
Patricia McQuilkin ◽  
Kanagasabai Udhayashankar ◽  
Eric Adu ◽  
Ann Moormann
Keyword(s):  
Treatment Outcomes ◽  
Severe Malaria ◽  
Medical Center ◽  
Laboratory Data ◽  
Case Fatality ◽  
Febrile Illness ◽  
Salmonella Typhi ◽  
Presenting Symptoms ◽  
National Referral Hospital ◽  
Children Under 5

Malaria is endemic in Liberia with a prevalence rate of up to 60% in some regions, and it has been a major cause of death in children under 5 years of age. Prior to the recent Ebola epidemic, we undertook a prospective, hospital-based pilot study at the National Referral Hospital in Monrovia, to characterize the presentation, accuracy of diagnosis, and treatment outcomes of children presenting for treatment of malaria. From June 2013 to May 2014, we recruited children 5 years and under who presented to the JFK Medical Center with suspected malaria. We collected both clinical and laboratory data on admission and on discharge. We enrolled 477 patients with an average age of 1.6 years. Demographic factors associated with testing negative for malaria included regular bed net use and prior treatment for malaria. The most common presenting symptoms of severe malaria in this population were headache and seizures. Of 246 patients admitted and treated for severe malaria, 33% tested negative by rapid diagnostic test and blood smear for malaria. The case fatality rate was higher for the patients who tested negative for malaria (4.9%) versus those who tested positive (0.6%). Three children who tested negative for malaria showed evidence of undiagnosed Salmonella typhi infection. These results suggest that malaria may be overdiagnosed and that the diagnoses of other infectious diseases, which present in a similar fashion, may be neglected. These findings underscore the need to develop rapid diagnostic tests to screen for alternative causes of febrile illness.

Rapid diagnosis of typhoid fever by PCR assay using one pair of primers from flagellin gene of Salmonella typhi

2003 ◽  
Vol 9 (3) ◽  
pp. 233-237 ◽  
Author(s):  
Muhammad Nasrum Massi ◽  
Akinobu Gotoh ◽  
Acharya Bishnu ◽  
Masato Kawabata ◽  
Toshiro Shirakawa ◽  
...  
Keyword(s):  
Typhoid Fever ◽  
Salmonella Typhi ◽  
Rapid Diagnosis ◽  
Pcr Assay ◽  
Flagellin Gene

scholarly journals Comparison and diagnosis of Entamoeba in stool sample from rural community of Nepal

2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Sandeep Thapa
Keyword(s):  
Stool Sample ◽  
Entamoeba Histolytica ◽  
Rural Community ◽  
Microscopic Examination ◽  
Nested Pcr ◽  
Health Knowledge ◽  
Intestinal Parasites ◽  
Intestinal Parasitosis ◽  
Microbiological Methods ◽  
Pcr Targeting

ObjectiveTo find out the prevalence of Entamoeba species in rural community of Nepal.The purpose of the study is to evaluate Nested PCR, microscopic examination and Elisa assay for detection and differentiation of Entamoeba species.IntroductionNepal being a developing country has many health problems, which escalate in intensity at different times of the year or in epidemic form.Amebiasis is one of the infectious diseases that is highly seen in rural area of Nepal caused by Entamoeba species ((1, 2). Recent reports show that open defecation, drinking untreated water, unsanitary habits and lack of basic health knowledge cause higher mortality and morbidity in our country.E. histolytica is an anaerobic pathogenic parasitic. However, E. dispar and E.moshkovskii exits as non-pathogenic. Likewise, E. histolytica, E. dispar and E. moshkovskii are morphologically identical but genetically distinct species (3).MethodsA total of 270 faecal sample were collected from south eastern terai region of Nepal after the informed consent form. The samples were processed by direct wet smear and formalin ethyl acetate concentration technique (4). Eventually, microscopic examination were performed for the detection of Entamoeba species along with other intestinal parasites. Furthermore, enzyme immunoassay were executed to detect antigens of E. histolytica through ELISA. Additionally, microscopically positive samples for Entamoeba species cysts were further characterized using a Nested- PCR targeting 16S-like ribosomal RNA gene (5). The PCR generate amplicons which was subjected to 2% agarose gels electrophoresis and visualized under UV transilluminator.Results8.52% of the total collected samples were microscopically positive for Entamoeba cysts either singly or in combination with other intestinal parasites. Likewise, among 270 stool sample, viral diarrheal was most significant form of diarrhoea found in 76.67% of patients. Among different organisms, As. Lumbricoids and E. histolytica, G. lambia and H. nana were identified in most of the patients accounting for 11.11%, 8.52%, 2.59% and 1.11% respectively. However, Lumbricoids, G. lambia, Tenia solium and E. histolytica were present in an individual patient while two patient was found with both As. Lumbricoids and G. lambia. Among several symptoms, diarrhoea seems to be the common symptoms infecting all of the patients which is followed by fever and vomiting which accounts for 55.1 % and 46.2% correspondingly. Whereas, nausea appears to be the least common symptoms infecting only 14.4% of patients.Subsequently, 56 cases were PCR positive, 51 cases were ELISA positive whereas 47 were found to be positive by microscopy.ConclusionsMolecular techniques are indeed promising tools for epidemiological studies, particularly in discriminating the pathogenic from the non-pathogenic species of the Entamoeba species. This study reports a new nested multiplex PCR strategy for detection and differentiation of E. histolytica, E. dispar and E. moshkovskii which is highly rapid, specific and sensitive which is useful for proper diagnosis, immunological assay and drug testing.References1. Chaudhary M, Maharjan M. ( 2014). Association of Anaemia with Parasitic Infection in Pregnant Women Attending Antenatal Clinic at Koshi Zonal Hospital. Neplalese Journal of Zoology, 2(1):1-7.2. Tandukar S, Ansari S, Adhikari N, Shrestha A, Gautam J, Sharma B, et al (2013). Intestinal parasitosis in school children of Lalitpur district of Nepal. BMC research notes, 6(1):449.3. Clark C, Espinosa Cantellano M, Bhattacharya A. Entamoeba histolytica: an overview of the biology of the organism (2013). Amebiasis: World Scientific, 1-45.4. Dhanabal J, Selvadoss PP, Muthuswamy K(2014). Comparative study of the prevalence of intestinal parasites in low socioeconomic areas from South Chennai, India. Journal of parasitology research.5. Van den Bossche, D., Cnops, L., Verschueren, J., & Van Esbroeck, M. (2015). Comparison of four rapid diagnostic tests, ELISA, microscopy and PCR for the detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in feces. Journal of microbiological methods, 110, 78-84.

scholarly journals Development of nested PCR based on the ViaB sequence to detect Salmonella typhi.

1995 ◽  
Vol 33 (11) ◽  
pp. 3082-3082 ◽  
Author(s):  
Y Hashimoto ◽  
Y Itho ◽  
Y Fujinaga ◽  
A Q Khan ◽  
F Sultana ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Salmonella Typhi

scholarly journals First report of Lihan Tick virus (Phlebovirus, Phenuiviridae) in ticks, Colombia

2020 ◽  
Author(s):  
Yesica Lopez ◽  
Jorge Miranda ◽  
Salim Mattar ◽  
Marco Gonzalez ◽  
Joel Rovnak
Keyword(s):  
Real Time ◽  
Nested Pcr ◽  
Rna Extraction ◽  
Febrile Illness ◽  
Domestic Animals ◽  
Positive Sample ◽  
Specific Primers ◽  
First Report ◽  
Clinical Syndromes ◽  
Disease Associations

Abstract Background: Tick-borne Phleboviruses (TBPVs) comprise human and animal viruses that can cause a variety of clinical syndromes ranging from self-limiting febrile illness to fatal haemorrhagic fevers.Objective: Detect Phlebovirus in ticks collected from domestic animals in Córdoba, Colombia.Methods: We collected 2,365 ticks from domestic animals in three municipalities of the Department of Cordoba, Colombia in 2016. Ticks were identified and pooled by species for RNA extraction. A nested real-time PCR with specific primers for Phlebovirus and a probe for Heartland virus (HRTV) were performed. Also, a conventional nested PCR, with the same specific primers was used to detect other Phleboviruses, with positive reactions indicated by an amplified cDNA fragment of approximately 244 bp determined by gel electrophoresis. These bands were gel-purified and sequenced by the Sanger method.Results: Using real-time RT-PCR, no positive results for HRTV were found. However, using conventional nested PCR 2.2% (5/229 pools) yielded a product of 244 bp. One positive sample was detected in a pool of Dermacentor nitens ticks collected from a horse, and the four remaining positive pools were from Rhipicephalus microplus collected from cattle. The five positive nucleotide sequences were identical to each other, with identities of 93% to 96% compared to a section of the L-segment of Lihan Tick virus, a Phlebovirus originally detected in R. microplus ticks in China. The strongest identity (96%-99%) was with Lihan Tick virus detected in R. microplus ticks from Brazil.Conclusions: This is the first report of viral detection in ticks in Colombia. We detected a Colombian strain of Lihan Tick virus. We recommend investigating possible disease associations with this tick-borne virus in Colombia.

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