New genus-specific primers for PCR identification of Rubrobacter strains

Abstract Toxoplasma gondii, a coccidian parasite known to infect almost all warm-blooded animals, is the cause of one of the most common zoonotic parasitic diseases. The aim of the study is to determine whether the 529 bp fragment or the TGR1E gene is more useful target for PCR identification of T. gondii, for common use. The brains of 221 carnivores and omnivores collected between 2013 and 2015 from north-eastern Poland were examined for the presence of this parasite. The DNA was extracted and then amplified using specific primers. Positive results were obtained in 24% of brain samples using the TGR1E target and 19% using the 529 bp sequence. The results demonstrate that both TGR1E and 529 bp repeat element are suitable for detecting T. gondii DNA in wildlife animals, and the combination of two methods is necessary to obtain reliable results.

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Morphological and molecular identification of Fusarium tricinctum and Fusarium acuminatum as causal agents of garlic bulbs rot in Serbia

Zbornik Matice srpske za prirodne nauke ◽

10.2298/zmspn1733271i ◽

2017 ◽

pp. 271-277 ◽

Cited By ~ 1

Author(s):

Maja Ignjatov ◽

Dragana Bjelic ◽

Zorica Nikolic ◽

Dragana Milosevic ◽

Jelena Marinkovic ◽

...

Keyword(s):

Molecular Identification ◽

Allium Sativum ◽

Fusarium Species ◽

Fungal Growth ◽

Specific Primers ◽

Fusarium Acuminatum ◽

The World ◽

Pcr Identification ◽

Fusarium Tricinctum ◽

Over Time

Garlic (Allium sativum L.) is considered to be one of the oldest crops in the world. During 2016, infected garlic bulbs occurred in storages on several localities of the Province of Vojvodina. Symptomatic cloves showed typical rot symptoms such as softened and spongy areas covered with white fungal growth with deep lesions formed on the cloves which became dry over time. A total of 36 isolates of Fusarium species were obtained from diseased cloves of garlic. Colony morphology and microscopic properties of isolated Fusarium species were recorded from the cultures grown on PDA and CLA, respectively. Identification of two chosen isolates was performed by sequencing the EF-1? gene. The TEF sequence of isolate JBL12 showed 100% similarity with several F. tricinctum sequences and sequence of JBL539 showed 99% identity with several F. acuminatum sequences and they were deposited in the NCBI GenBank. Based on the results of the morphological and molecular identification, isolates JBL12 and JBL539 were identified as F. tricinctum and F. acuminatum, respectively, as new causal agents of garlic bulbs rot in Serbia. Specific primers were designed for the PCR identification of the F. tricinctum.

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Determination of Helicobacter pylori Virulence Genes in Gastric Biopsies by PCR

ISRN Gastroenterology ◽

10.1155/2013/606258 ◽

2013 ◽

Vol 2013 ◽

pp. 1-4 ◽

Cited By ~ 8

Author(s):

Tamer Essawi ◽

Wail Hammoudeh ◽

Israr Sabri ◽

Walid Sweidan ◽

Mohammad A. Farraj

Keyword(s):

Helicobacter Pylori ◽

Virulence Genes ◽

Gastric Biopsy ◽

Strong Indication ◽

Specific Primers ◽

Biopsy Specimens ◽

Pcr Identification ◽

Gastric Biopsies ◽

H Pylori

Aim. The aim of this study was to identify the presence of H. pylori in biopsy specimens from symptomatic patients by PCR. In addition, the rate of cagA, vacA, iceA1, and iceA2 virulence genes was determined. Materials and Methods. One hundred antral gastric biopsy specimens were collected during endoscopy from patients suffering from gastroduodenal symptoms. The samples were collected by the gastroenterologists in their own clinics in Ramallah, Palestine. DNA was extracted from the biopsies and subsequently used for PCR identification of H. pylori and the virulence genes using specific primers. Results. The rate of positive H. pylori in the collected biopsies was 44%. The rates of the virulence genes in this sample: cagA, vacA, iceA1, and iceA2 were 65.9%, 40.9%, 63.6%, and 84.1%, respectively. Conclusion. The iceA2 gene was the most frequent in this study. Much research is necessary to determine the presence of an association of this gene with gastric pathology. Variation in the rates of the iceA gene in different countries is a strong indication of its geographical distribution. This study would provide important information regarding the prevalence of virulence genes (vacA, cagA, iceA1, and iceA2) in H. pylori strains in the sample tested in this country.

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PCR amplification of a putative gene for exo-1,3-β-glucanase to identify the pathogenic oomycete Pythium insidiosum

Asian Biomedicine ◽

10.5372/1905-7415.0805.338 ◽

2014 ◽

Vol 8 (5) ◽

pp. 637-644 ◽

Cited By ~ 12

Author(s):

Angsana Keeratijarut ◽

Tassanee Lohnoo ◽

Wanta Yingyong ◽

Umporn Nampoon ◽

Tassanee Lerksuthirat ◽

...

Keyword(s):

Diagnostic Performance ◽

Disease Diagnosis ◽

Detection Sensitivity ◽

Molecular Diagnostic ◽

Molecular Diagnostic Techniques ◽

Specific Primers ◽

Pythium Insidiosum ◽

Putative Gene ◽

Pcr Identification ◽

Primer Sets

AbstractBackground: Pythium insidiosum is the etiologic agent of pythiosis, a life-threatening infectious disease. Diagnosis of pythiosis is difficult and often delayed. Early diagnosis can lead to prompt treatment, and therefore a better prognosis for patients with pythiosis. Molecular diagnostic techniques are useful if microbiological and immunological assays are not available, or in cases of suspected pythiosis that test negative by other methods. So far, PCR identification of P. insidiosum has been largely relied on amplification of the rDNA region.Objective: To evaluate the diagnostic performance of Dx3 and Dx4 primers specific for a putative gene for exo- 1,3-β-glucanase (PinsEXO1), which encodes a specific immunogen of P. insidiosum, for rapid single-round PCR identification of P. insidiosum, in comparison with the previously-reported rDNA-specific primers, ITSpy1 and ITSpy2.Materials and Methods: Genomic DNA (gDNA) from 35 P. insidiosum isolates and 48 control organisms were prepared to evaluate the diagnostic performance of the PinsEXO1- and rDNA-specific primers.Results: When amplifying the control gDNA by using the Dx3/4 and ITSpy1/2 primer sets, no PCR product was observed, indicating that both primer sets had 100% detection specificity. When amplifying the P. insidiosum gDNA, the Dx3/4 primers provided an expected 550-bp amplicon for all 35 isolates, while the ITSpy1/2 primers provided an expected 230-bp amplicon for only 32 isolates. Thus, detection sensitivity of the Dx3/4 and ITSpy1/2 primer sets were 100% and 91%, respectively.Conclusion: By using the Dx3/4 primers, PinsEXO1 was an alternative, efficient, and novel PCR target for rapid single-round PCR identification of P. insidiosum.

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