scholarly journals Leptospiral Culture without 5’-Fluorouracil Revealed Improved Leptospira Isolation from Febrile Patients in North-Eastern Malaysia

2020 ◽  
Vol 17 (4) ◽  
pp. 1307
Author(s):  
Amira Wahida Mohamad Safiee ◽  
Mohammad Ridhuan Mohd Ali ◽  
Mohd Hashairi Fauzi ◽  
Alwi Muhd Besari ◽  
Chan Yean Yean ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Whole Blood ◽  
Rapid Test ◽  
Febrile Illness ◽  
Culture Method ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Leptospira Interrogans ◽  
Acute Febrile Illness ◽  
Leptospira Spp

Objectives: Isolation of Leptospira by culture represents a definitive growth and confirmation of the disease, yet it is hampered with its nature of slow growth. With slight modification of culture method, the study aims to isolate and characterize Leptospira spp. from patients with acute febrile illness. Methods: A total of 109 blood samples were collected from patients with acute febrile illness that presented at the Emergency Department of Hospital Universiti Sains Malaysia, Malaysia. Clinical samples were subjected to Leptospira IgM Rapid test, microscopic agglutination test (MAT), isolation by culture method, and direct real-time PCR test. For leptospiral isolation, the samples (whole blood and deposit from spun plasma) were cultured into modified Ellinghausen McCullough Johnson Harris (EMJH) media with and without 5’-fluorouracil (5-FU). In every culture positive sample, partial 16S rRNA gene sequencing was performed for molecular identification of the isolates. Phylogenetic analysis was carried out to determine the genetic relatedness among the isolates. An inhibition of 5-FU study was performed on Leptospira interrogans serovar Canicola with different concentrations to compare the growth detection of the tested Leptospira with or without 5-FU within 7 days of incubation. Results: Leptospirosis was diagnosed in 14.7% of patients with acute febrile illness. Two Leptospira spp. (n = 2/109, 1.85%) were successfully isolated from whole blood and deposit from spun plasma samples. B004 and B208 samples were positive at day 11 and day 7, respectively, in EMJH media without addition of 5-FU. Sample B004 was identified as Leptospira interrogans and B208 as Leptospira weilli. Phylogenetic analysis confirmed that both of them were within pathogenic group and they were not related. The 5-FU inhibition study revealed that additional of 5-FU at final concentration of 200 µg/mL to EMJH media demonstrated an inhibitory effect on the growth of the tested strain Conclusion: Isolation of Leptospira spp. using EMJH media without addition of 5’-fluorouracil resulted in a better outcome. Two pathogenic Leptospira isolates were successfully cultivated from patients with acute febrile illness that were genetically not related.

scholarly journals Detection of NS1 antigen, IgM and IgG antibodies using a commercial Dengue rapid test kit for the diagnosis of dengue infection in patients with acute febrile illness

2021 ◽  
Vol 8 (3) ◽  
pp. 235-238
Author(s):  
Nalamanda Suma ◽  
T Sarada
Keyword(s):  
Solid Phase ◽  
Serological Test ◽  
Dengue Infection ◽  
Rapid Test ◽  
Febrile Illness ◽  
Acute Febrile Illness ◽  
Igg Antibodies ◽  
Igm Antibody ◽  
Test Kit ◽  
Ns1 Antigen

Dengue is an endemic arboviral illness. With the increasing incidence of dengue infection, an early diagnostic confirmation of dengue infection in patients facilitates timely clinical intervention, etiological investigation, and disease control. Objective of this study was to evaluate a commercially available serological test kit - Dengue Day 1 Test. This is for the detection of dengue NS1 antigen, differential detection of IgM and IgG antibodies on a single acute serum sample. Atotal of 100 patients with acute febrile illness were included in this study. Serum samples were analysed for Dengue NS1 Ag, IgM & IgG Antibodies using a commercially available Dengue Day 1 Test, rapid solid phase immuno-chromatographic test.As many as 23 (23%) samples were NS1 positive and 17 (17%) samples were positive for IgM antibodies. Based on the combination of dengue NS1 antigen and IgM antibody test, total of 34 patients (34%) were positive for dengue virus infection.Results of the study suggested that a combination of dengue NS1 antigen and IgM antibody tests would increase the rate of detection of dengue illness. This combination would increase the efficacy and aid in early diagnosis of dengue infection.

scholarly journals Rapid-Test Based Identification of Influenza as an Etiology of Acute Febrile Illness in Cambodia

2011 ◽  
Vol 85 (6) ◽  
pp. 1144-1145 ◽  
Author(s):  
Matthew R. Kasper ◽  
Ly Sovann ◽  
Thomas F. Wierzba ◽  
Chadwick Y. Yasuda ◽  
Shannon D. Putnam ◽  
...  
Keyword(s):  
Rapid Test ◽  
Febrile Illness ◽  
Acute Febrile Illness

scholarly journals Characterization of clinical Ralstonia strains and their taxonomic position

2021 ◽  
Author(s):  
Ad C. Fluit ◽  
Jumamurat R. Bayjanov ◽  
María Díez Aguilar ◽  
Rafael Cantón ◽  
Michael M. Tunney ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Virulence Factors ◽  
Ralstonia Solanacearum ◽  
Novel Species ◽  
Species Group ◽  
Clinical Samples ◽  
Rrna Gene ◽  
Minimal Inhibitory Concentrations ◽  
Group A ◽  
Group B

AbstractTo improve understanding of the role of Ralstonia in cystic fibrosis (CF), whole genomes of 18 strains from clinical samples were sequenced using Illumina technology. Sequences were analysed by core genome Multi-Locus Sequence Typing, Average Nucleotide Identity based on BLAST (ANIb), RAST annotation, and by ResFinder. Phylogenetic analysis was performed for the 16S rRNA gene, and the OXA-22 and OXA-60 ß-lactamase families. The minimal inhibitory concentrations (MICs) were determined using broth microdilution. ANIb data for the 18 isolates and 54 strains from GenBank, supported by phylogenetic analysis, showed that 8 groups of clusters (A-H), as well as subgroups that should be considered as species or subspecies. Groups A-C contain strains previously identified as Ralstonia solanacearum and Ralstonia pseudosolanacearum. We propose that group A is a novel species. Group B and C are Ralstonia syzygii, Ralstonia solanacearum, respectively. Group D is composed of Ralstonia mannitolilytica and Group E of Ralstonia pickettii. Group F and G should be considered novel species. Group H strains belong to R. insidiosa. OXA-22 and OXA-60 family ß-lactamases were encoded by all strains. Co-trimoxazole generally showed high activity with low MICs (≤1 mg/l) as did ciprofloxacin (≤0.12 mg/l). MICs against the other antibiotics were more variable, but generally high. RAST annotation revealed limited differences between the strains, and virulence factors were not identified. The taxonomy of the genus Ralstonia is in need of revision, but sequencing additional isolates is needed. Antibiotic resistance levels are high. Annotation did not identify potential virulence factors.

scholarly journals Comparison between latex and microscopic agglutination test for detection of human leptospiral antibodies

2016 ◽  
Vol 10 (3) ◽  
pp. 219
Author(s):  
Ehsanollah Sakhaee ◽  
Mehdi Golchin ◽  
Zahra Davoodian
Keyword(s):  
Febrile Illness ◽  
High Sensitivity ◽  
Latex Agglutination ◽  
Agglutination Test ◽  
Basic Method ◽  
Microscopic Agglutination Test ◽  
Leptospira Interrogans ◽  
Acute Febrile Illness ◽  
Specific Antibodies ◽  
Human Leptospirosis

Leptospirosis, notably Weil’s syndrome, is an often severe, acute febrile illness caused by microorganisms of the genus <em>Leptospira</em>. The microscopic agglutination test (MAT) is the basic method for the sero-diagnosis of leptospirosis, as the test has a high sensitivity and can be used for classification, but has some disadvantages. Therefore, we have explored latex agglutination test (LAT) for use as a practical and rapid for sero-diagnosis of human leptospirosis and compared the applicability of commercial tests MAT and LAT in the detection of specific antibodies against <em>Leptospira interrogans</em> in human.

scholarly journals Molecular Confirmation of Co-Infection by Pathogenic Leptospira spp. and Orientia tsutsugamushi in Patients with Acute Febrile Illness in Thailand

2013 ◽  
Vol 89 (4) ◽  
pp. 797-799 ◽  
Author(s):  
Piengchan Sonthayanon ◽  
Wirongrong Chierakul ◽  
Vanaporn Wuthiekanun ◽  
Direk Limmathurotsakul ◽  
Premjit Amornchai ◽  
...  
Keyword(s):  
Febrile Illness ◽  
Acute Febrile Illness ◽  
Orientia Tsutsugamushi ◽  
Pathogenic Leptospira ◽  
Leptospira Spp

scholarly journals Molecular Characterisation and Phylogenetic Analysis of Dermatophytic Fungi Isolated from Tinea Capitis in Northwest Nigeria Using Sequence of the 28S rRNA

2021 ◽  
Vol 12 (3) ◽  
pp. 646-655
Author(s):  
Hussain Yahaya Ungo-kore ◽  
Joseph Olorunmola Ehinmidu ◽  
Josiah Ademola Onaolapo ◽  
Olayeni Stephen Olonitola
Keyword(s):  
Phylogenetic Analysis ◽  
Tinea Capitis ◽  
28S Rdna ◽  
Clinical Samples ◽  
28S Rrna ◽  
Trichophyton Tonsurans ◽  
Detection And Identification ◽  
Microsporum Audouinii ◽  
Trichophyton Simii ◽  
Dermatophyte Species

The detection and identification of fungal DNA from clinical samples is one of the fundamental approaches in biomedicine. The incidence, distribution, and control of dermatophytes has progress significantly and the use of phylogenetic species concepts based on rRNA regions have enhanced the taxonomy of dermatophyte species; however, the use of 28S rDNA genes has certain limitations. This gene has been used in dermatophyte taxonomy with limited enumeration; we appraised the sequence disparity within and among groups of the species, the gene ranking in identification, phylogenetic analysis, and taxonomy of 32 strains of eight dermatophyte species. In this study, a set of primers was adopted to amplify the target followed by a partial sequencing of the rDNA. The utilization of a pairwise nucleotide differentiation, an affinity was observed among eight dermatophyte species, with disparity among species ranging from 0 to 197 base pair (bp). Intra-species bp differences were found within strains of Trichophyton eriotrephon, Trichophyton bullosum, Trichophyton simii (Trichophyton genus), Microsporum audouinii, and Trichophyton tonsurans (Microsporum and Trichophyton genus, respectively); however, only some strains of Trichophyton eriotrephon were found to be invariant having three genotypes. Trichophyton tonsurans exhibited most intra-species variability. The characterization and construction of a phylogenetic tree of 28S rDNA gene on dermatophyte species provide a bedrock of an additional finding of connections between species. However, 28S rRNA capture provides a novel method of effective and sensitive detection of dermatophytes lodged in human skin scale. We report for the first time the emergence of T. eriotrephon, T. bullosum, T. simii, T. benhamiae, and Ctenomyces serratus dermatophytes from Tinea capitis in Nigeria.

scholarly journals First Molecular Confirmation of Equine Ocular Setaria digitata in China

2021 ◽  
Vol 8 (4) ◽  
pp. 55
Author(s):  
Feng Yu ◽  
Bo Liu ◽  
Shulei Chen ◽  
Ziwen Yi ◽  
Xianyong Liu ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Light Microscopy ◽  
Corneal Opacity ◽  
12S Rrna ◽  
Rrna Gene ◽  
Asian Countries ◽  
Setaria Digitata ◽  
12S Rrna Gene ◽  
The Right ◽  
Ocular Discharge

A 5-year-old Mongolian mare (Equus caballus Linnaeus, 1758) was observed to have corneal opacity and excessive ocular discharge. An ophthalmic examination revealed a moving thread-like cylindrical worm in the anterior chamber of the right eye. The parasite was successfully removed surgically. The worm was observed under light microscopy and confirmed as Setaria digitata by 12S rRNA gene amplification and sequencing. Phylogenetic analysis demonstrated similarity with Setaria digitata in the National Center for Biotechnology Information (NCBI) GenBank database isolated from other Asian countries. This report is the first confirmed case of equine ocular setariasis by molecular diagnosis in China, which may indicate its presence in livestock and promote research on its epidemiology.

scholarly journals Assessment of Genetic Diversity and Symbiotic Efficiency of Selected Rhizobia Strains Nodulating Lentil (Lens culinaris Medik.)

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 15
Author(s):  
Badreddine Sijilmassi ◽  
Abdelkarim Filali-Maltouf ◽  
Hassan Boulahyaoui ◽  
Aymane Kricha ◽  
Kenza Boubekri ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Phylogenetic Analysis ◽  
16S Rrna ◽  
Rhizobium Leguminosarum ◽  
Sequence Similarity ◽  
P Value ◽  
Rrna Gene ◽  
Research Station ◽  
Symbiotic Efficiency ◽  
Rhizobium Strains

A total of 14 Rhizobium strains were isolated from lentil accessions grown at the ICARDA experimental research station at Marchouch in Morocco and used for molecular characterization and symbiotic efficiency assessment. Individual phylogenetic analysis using the 16S rRNA gene, house-keeping genes rpoB, recA, and gyrB, and symbiotic genes nodD and nodA along with Multilocus Sequence Analysis (MLSA) of the concatenated genes (16S rRNA-rpoB-recA-gyrB) was carried out for the identification and clustering of the isolates. The symbiotic efficiency of the strains was assessed on three Moroccan lentil cultivars (Bakria, Chakkouf, and Zaria) based on the number of nodules, plant height, plant dry weight, and total nitrogen content in leaves. The results showed that the individual phylogenetic analysis clustered all the strains into Rhizobium laguerreae and Rhizobium leguminosarum with sequence similarity ranging from 94 to 100%, except one strain which clustered with Mesorhizobium huakuii with sequence similarity of 100%. The MLSA of the concatenated genes and the related percentages of similarity clustered these strains into two groups of Rhizobium species, with one strain as a new genospecies when applying the threshold of 96%. For symbiotic efficiency, the Bakria variety showed the best association with 10 strains compared to its non-inoculated control (p-value ≤ 0.05), followed by Chakkouf and Zaria. The present study concluded that the genetic diversity and the symbiotic efficiency of Rhizobium strains appeared to be mainly under the control of the lentil genotypes.

Using a Polymerase Chain Reaction as a Complementary Test to Improve the Detection of Dermatophyte Fungus in Nails

2014 ◽  
Vol 104 (3) ◽  
pp. 233-237 ◽  
Author(s):  
María José Iglesias Sánchez ◽  
Ana María Pérez Pico ◽  
Félix Marcos Tejedor ◽  
María Jesús Iglesias Sánchez ◽  
Raquel Mayordomo Acevedo
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Classical Method ◽  
Clinical Manifestations ◽  
Culture Method ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Traditional Procedure ◽  
Polymerase Chain

Background Dermatomycoses are a group of pathologic abnormalities frequently seen in clinical practice, and their prevalence has increased in recent decades. Diagnostic confirmation of mycotic infection in nails is essential because there are several pathologic conditions with similar clinical manifestations. The classical method for confirming the presence of fungus in nail is microbiological culture and the identification of morphological structures by microscopy. Methods We devised a nested polymerase chain reaction (PCR) that amplifies specific DNA sequences of dermatophyte fungus that is notably faster than the 3 to 4 weeks that the traditional procedure takes. We compared this new technique and the conventional plate culture method in 225 nail samples. The results were subjected to statistical analysis. Results We found concordance in 78.2% of the samples analyzed by the two methods and increased sensitivity when simultaneously using the two methods to analyze clinical samples. Now we can confirm the presence of dermatophyte fungus in most of the positive samples in just 24 hours, and we have to wait for the result of culture only in negative PCR cases. Conclusions Although this PCR cannot, at present, substitute for the traditional culture method in the detection of dermatophyte infection of the nails, it can be used as a complementary technique because its main advantage lies in the significant reduction of time used for diagnosis, in addition to higher sensitivity.

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