Estimating the prevalence of mixed-type gonococcal infections in Queensland, Australia

Sexual Health ◽  
2015 ◽  
Vol 12 (5) ◽  
pp. 439 ◽  
Author(s):  
Ella Trembizki ◽  
Christine Doyle ◽  
Cameron Buckley ◽  
Amy Jennison ◽  
Helen Smith ◽  
...  
Keyword(s):  
Sequence Data ◽  
Nucleic Acid Amplification ◽  
Clinical Samples ◽  
Anatomical Site ◽  
Diagnostic Screening ◽  
Specific Pcr ◽  
Screening Population ◽  
Pcr Products ◽  
Polymerase Chain ◽  
The One

Background Mixed gonococcal infections within the one anatomical site have been recognised but questions remain over how often they occur. In this study, the aim was to estimate the prevalence of mixed gonococcal infections using novel real-time polymerase chain reaction (PCR) methods that were developed and validated, targeting the gonococcal porB gene. Methods: Neisseria gonorrhoeae strains were categorised into three different porB groups, based on sequence data derived from N. gonorrhoeae multi-antigen sequence typing (NG-MAST) analyses of local isolates. Specific PCR methods for each group were then developed and these PCR methods were used to test clinical samples (n = 350) that were positive for gonorrhoea as determined by nucleic acid amplification test (NAAT) diagnostic screening. Results: Initial validation using isolates showed the group PCR methods proved 100% sensitive and 100% specific for their respective porB groups. When applied to the clinical specimens, 298/350 (85%) provided positive results by the group PCR methods. Of these, four specimens showed evidence of mixed infections, supported by subsequent DNA sequencing of the PCR products. Conclusions: The data provide further evidence of mixed gonococcal infections at the same anatomical site, but show that such infections may be relatively infrequent (1.3%; 95% confidence interval 0.01–2.6%) in a general screening population.

scholarly journals Characterization of a New Alloantigen (SH) on the Human Neutrophil Fcγreceptor IIIb

Blood ◽  
1997 ◽  
Vol 89 (3) ◽  
pp. 1027-1034 ◽  
Author(s):  
Juergen Bux ◽  
Ernst-Ludwig Stein ◽  
Philippe Bierling ◽  
Patricia Fromont ◽  
Mary Clay ◽  
...  
Keyword(s):  
Nucleotide Sequence Analysis ◽  
Polymorphism Analysis ◽  
Mutational Event ◽  
Coding Region ◽  
Specific Pcr ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Antigen Frequency ◽  
Neonatal Neutropenia

Abstract Polymorphic structures of the neutrophil Fcγreceptor IIIb (FcγRIIIb) result in alloantibody formation that causes alloimmune neonatal neutropenia and transfusion reactions. Alloantigens located on FcγRIIIb include the antigens NA1 and NA2. In four cases of alloimmune neonatal neutropenia, granulocyte-specific alloantibodies directed against a thus far unknown antigen were detected by granulocyte agglutination and immunofluorescence tests in the maternal sera. By the use of the monoclonal antibody–specific immobilization of granulocyte antigens (MAIGA) assay, the new antigen, termed SH, was located on the FcγRIIIb. Nucleotide sequence analysis of the FcγRIIIb coding region from a SH(+) individual showed a single-base C→A mutation at position 266, which results in an Ala78Asp amino acid substitution. A family study confirmed that this nucleotide difference is inherited, and corresponds to the SH phenotype. Serologic typing of 309 randomly selected individuals showed an antigen frequency of 5% in the white population. The same frequency was found by genotyping, for which a technique based on polymerase chain reaction (PCR) using sequence-specific primers (PCR-SSP) was developed. Typing of all SH(+) individuals for NA1 and NA2, and PCR-restriction fragment length polymorphism analysis of the NA-specific PCR products from five SH(+) individuals using the SH-specific endonuclease SfaN I showed that SH antigen is very probably the result of an additional mutational event in the NA2 form of the FcγRIIIB gene. Immunochemical studies also demonstrated that the SH determinants reside on the 65- to 80-kD NA2 isoform of the FcγRIIIb. Our findings show the existence of an additional polymorphism of the FcγRIIIb, which can result in alloantibody formation causing alloimmune neonatal neutropenia.

scholarly journals Spread of TEM, VIM, SHV, and CTX-Mβ-Lactamases in Imipenem-Resistant Gram-Negative Bacilli Isolated from Egyptian Hospitals

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
El sayed Hamdy Mohammed ◽  
Ahmed Elsadek Fakhr ◽  
Hanan Mohammed El sayed ◽  
Said abd Elmohsen Al Johery ◽  
Wesam Abdel Ghani Hassanein
Keyword(s):  
Direct Sequencing ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Therapeutic Problem

Carbapenem-resistant Gram-negative bacilli resulting fromβ-lactamases have been reported to be an important cause of nosocomial infections and are a critical therapeutic problem worldwide. This study aimed to describe the prevalence of imipenem-resistant Gram-negative bacilli isolates and detection ofblaVIM,blaTEM,blaSHV,blaCTX-M-1, andblaCTX-M-9genes in these clinical isolates in Egyptian hospitals. The isolates were collected from various clinical samples, identified by conventional methods and confirmed by API 20E. Antibiotic susceptibility testing was determined by Kirby-Bauer technique and interpreted according to CLSI. Production ofblaVIM,blaTEM,blaSHV, andblaCTX-Mgenes was done by polymerase chain reaction (PCR). Direct sequencing from PCR products was subsequently carried out to identify and confirm theseβ-lactamases genes. Out of 65 isolates, (46.1%) Escherichia coli, (26.2%) Klebsiella pneumoniae, and (10.7%) Pseudomonas aeruginosa were identified as the commonest Gram-negative bacilli. 33(50.8%) were imipenem-resistant isolates. 22 isolates (66.7%) carriedblaVIM, 24(72.7%) hadblaTEM, and 5(15%) showedblaSHV, while 12(36%), 6(18.2%), and 0(0.00%) harboredblaCTX-M-1,blaCTX-M-9, andblaCTX-M-8/25, respectively. There is a high occurrence ofβ-lactamase genes in clinical isolates and sequence analysis of amplified genes showed differences between multiple SNPs (single nucleotide polymorphism) sites in the same gene among local isolates in relation to published sequences.

scholarly journals Comparison of Two Multiplex PCR Systems for Meat Species Authentication

2019 ◽  
Author(s):  
D. Al-taghlubee ◽  
A. Misaghi ◽  
P. Shayan ◽  
A. Akhondzadeh Basti ◽  
H. Gandomi ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Cross Reactivity ◽  
Specific Pcr ◽  
Pcr Products ◽  
Reverse Primer ◽  
Specific Reverse Primer ◽  
Meat Species ◽  
Polymerase Chain ◽  
Species Authentication ◽  
Species Specific

Background: Meat species adulteration has become a problem of concern. This study aimed to compare two previously published multiplex Polymerase Chain Reaction (PCR) methods for meat species authentication.  Methods: The primers used in the first multiplex PCR involved species-specific reverse primer for sheep, goat, cattle, pig, and donkey with universal forward primer. In the second multiplex PCR, the primers included species-specific forward and reverse primer for pork, lamb, ostrich, horse, and cow. The extracted DNA was then amplified with species-specific primers and with mix primers separately in the respective multiplex PCR. Results: The first multiplex PCR was accompanied with cross reactivity, whereas the second multiplex PCR was specific as expected for pork, lamb, ostrich, horse, and cow. The first set of multiplex PCR showed not always amplification of all species-specific DNAs with a mixture of DNA from mentioned animals. Regarding the second set of primers, the extracted DNA of different meat species was amplified with corresponding species primers as simplex PCR resulting in specific amplicons for species DNA prepared from sheep, ostrich, horse, pig, and cattle with the specific PCR products of 119, 155, 253, 100, and 311 bp, respectively. Conclusion: Based on the present investigation, we recommend the multiplex PCR with the second set of primers included species-specific forward and reverse primers for species authentication of five meat types, including pork, lamb, ostrich, horse, as well as cow.

The development of PCR methodology for the identification of species of the tapeworm Moniezia from cattle, goats and sheep in central Vietnam

2011 ◽  
Vol 86 (4) ◽  
pp. 426-429 ◽  
Author(s):  
T.D. Nguyen ◽  
Q.D. Le ◽  
V.V. Huynh ◽  
S.T. Nguyen ◽  
T.V. Nguyen ◽  
...  
Keyword(s):  
Domestic Animals ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Domestic Ruminants ◽  
Central Vietnam ◽  
5.8S Rrna ◽  
Pcr Products ◽  
Identification Of Species ◽  
Polymerase Chain

AbstractThe aims of this study were to investigate the prevalence of Moniezia spp. in domestic ruminants in central Vietnam and to develop a polymerase chain reaction (PCR) technique to distinguish M. expansa from M. benedeni. Among 2040 examined domestic animals (540 cattle, 800 goats, 700 sheep) Moniezia was recovered from 5.4% of cattle, 16.4% of sheep and 20.6% of goats. A set of primers for PCR was designed to classify M. expansa and M. benedeni based on the amplification of DNA corresponding to the internal transcribed spacer of 5.8S rRNA. The 457 specimens (75 from cattle, 162 from goats, 150 from sheep, 30 from horses, 30 from chickens and 10 from dogs) were subjected to PCR for classification of Moniezia spp. PCR products with the expected sizes were amplified from bovine, ovine and caprine specimens. No specific PCR products were found for specimens from horses, chickens and dogs. Of the 75 specimens from cattle, nine were classified as M. expansa and 66 were M. benedeni. Among 162 caprine specimens, 138 were M. expansa and 24 were M. benedeni. The distribution of M. expansa and M. benedeni in 150 ovine specimens was 132 and 18, respectively. These results show that M. expansa is dominant in goats and sheep, whereas M. benedeni is more common in cattle; PCR can be used for classification of these two species.

scholarly journals Prevalence of Campylobacter spp. among imported broiler drumsticks and wings sold in Lithuania

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Dominyka Baltutytė ◽  
Laura Babonytė ◽  
Sigita Ramonaitė
Keyword(s):  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Chain Reaction ◽  
Microbiological Methods ◽  
Pcr Products ◽  
Pcr Method ◽  
Polymerase Chain ◽  
One Year ◽  
The One ◽  
Campylobacter Spp

The aim of this research was to estimate the prevalence of Campylobacter in imported broiler drumsticks and wings. During the one-year study period, 138 imported broiler samples (raw wings and drumsticks) were collected and tested from 3 different sellers. Campylobacter spp. were detected and isolated using traditional microbiological methods, identified using a multiplex polymerase chain reaction (PCR) method. The results of PCR products were analysed in agarose gel using electrophoresis. After an epidemiological study, C. jejuni and C. coli strains were selected and the prevalence of virulence genes was evaluated. The study identified Campylobacter spp. in 36 (26.1%) samples – 19 raw wings (27.9%) and 17 raw drumsticks (24.3%) samples were infected with these bacteria. Campylobacter spp. were most frequently detected in raw broiler samples during autumn (September–November) (47.2%) and winter (December–February) (41.6%) periods than spring (March–May) (5.5%) or summer (June–August) (5.5%). Contamination of products was not significantly impacted by the sale location (p > 0.05). The examination of virulence factors of Campylobacter spp. revealed that C. jejuni and C. coli strains contain 2 out of 3 virulence genes – CadF and CdtA. The CdtA gene was found in nearly all tested Campylobacter spp. strains isolated from broiler samples (94.4%).

scholarly journals Detection of Bovine Group B Rotaviruses in Feces by Polymerase Chain Reaction

1994 ◽  
Vol 6 (3) ◽  
pp. 302-307 ◽  
Author(s):  
Jarasvech Chinsangaram ◽  
Geoffrey Y. Akita ◽  
Bennie I. Osburn
Keyword(s):  
Polymerase Chain Reaction ◽  
Sequence Data ◽  
Genome Segment ◽  
Chain Reaction ◽  
Bovine Rotavirus ◽  
Detection Assay ◽  
Pcr Products ◽  
Group A ◽  
Polymerase Chain ◽  
Group B

A pair of primers designed from the sequence of genome segment 9 of group B rat rotavirus (IDIR) were employed to amplify genome segment 9 of a group B bovine rotavirus in a polymerase chain reaction (PCR) and to sequence the derived PCR products. A new pair of primers were synthesized from the obtained sequence data and used in a PCR detection assay for group B bovine rotavirus in fecal samples. In addition, another pair of primers were designed to produce a PCR-derived internal probe. This probe was used in a chemiluminescent hybridization to confirm the specificity and to increase the sensitivity of the assay. This assay could detect 0.1 fg of target double-stranded RNA. It was specific to group B bovine rotavirus and did not detect group B rat (IDIR) and porcine rotaviruses, group A bovine (NCDV), simian (SA-11), equine (H-2), porcine (OSU), human (DS-1), deer, and avian rotaviruses, coronavirus, or other enteric organisms tested in this study.

Analysis of the inhibition of nucleic acid dyes on polymerase chain reaction by capillary electrophoresis

2016 ◽  
Vol 8 (11) ◽  
pp. 2330-2334 ◽  
Author(s):  
Zhenqing Li ◽  
Chenchen Liu ◽  
Siyao Ma ◽  
Dawei Zhang ◽  
Yoshinori Yamaguchi
Keyword(s):  
Polymerase Chain Reaction ◽  
Capillary Electrophoresis ◽  
Nucleic Acid ◽  
Acid Dyes ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Pcr Product ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Accurate Quantification

An integrated polymerase chain reaction (PCR) and capillary electrophoresis (CE) system can realize accurate quantification of the target PCR product by adding labeling dyes to the PCR reagents, because CE can discriminate all the subsequent nucleic acids, including the primers, non-specific and specific PCR products.

scholarly journals Rapid and Sensitive Detection of Salmonella spp. Using CRISPR-Cas13a Combined With Recombinase Polymerase Amplification

2021 ◽  
Vol 12 ◽  
Author(s):  
Bailin An ◽  
Hongbin Zhang ◽  
Xuan Su ◽  
Yue Guo ◽  
Tao Wu ◽  
...  
Keyword(s):  
Real Time ◽  
Detection Method ◽  
Reaction System ◽  
Recombinase Polymerase Amplification ◽  
Clinical Samples ◽  
Specific Detection ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Polymerase Chain ◽  
The One

Salmonella spp. is one of the most common foodborne disease-causing pathogens that can cause severe diseases in very low infectious doses. Rapid and sensitive detecting Salmonella spp. is advantageous to the control of its spread. In this study, a conserved short fragment of the Salmonella invA gene was selected and used to design primers and specific crRNA (CRISPR RNA) for establishing a one-tube and two-step reaction system for Salmonella spp. detection, by combining recombinase polymerase amplification (RPA) with CRISPR-Cas13a (Clustered Regularly Interspaced Short Palindromic Repeats associated protein 13a) cleavage. The established one-tube RPA-Cas13a method can complete the detection within 20 min and the two-step RPA-Cas13a method detection time within 45 min. The designed primers were highly specific to Salmonella spp. and had no cross-reaction with the other nine diarrheal bacteria. The one-tube RPA-Cas13a could detect the Salmonella genome with the limit of 102 copies, which was the same as real-time polymerase chain reaction (PCR), but less sensitive than two-step RPA-Cas13a (100 copies). The detection results of one-tube or two-step RPA-Cas13a and real-time PCR were highly consistent in clinical samples. One-tube RPA-Cas13a developed in this study provides a simple, rapid, and specific detection method for Salmonella spp. While two-step assay was more sensitive and suitable for samples at low abundance.

scholarly journals Effectiveness and Durability of the Rice Pi-ta Gene in Yunnan Province of China

2014 ◽  
Vol 104 (7) ◽  
pp. 762-768 ◽  
Author(s):  
Jinbin Li ◽  
Lin Lu ◽  
Yulin Jia ◽  
Chengyun Li
Keyword(s):  
Dna Sequence ◽  
Yunnan Province ◽  
Sequence Variation ◽  
Pcr Amplification ◽  
Base Substitution ◽  
Field Isolates ◽  
Specific Pcr ◽  
Dna Sequence Variation ◽  
Pcr Products ◽  
Polymerase Chain

Rice blast is one of the most damaging diseases of rice worldwide. In the present study, we analyzed DNA sequence variation of avirulence (AVR) genes of AVR-Pita1 in field isolates of Magnaporthe oryzae in order to understand the effectiveness of the resistance gene Pi-ta in China. Genomic DNA of 366 isolates of M. oryzae collected from Yunnan province of China were used for polymerase chain reaction (PCR) amplification to examine the existence of AVR-Pita1 using gene-specific PCR markers. Results of PCR products revealed that 218 isolates of M. oryzae carry AVR-Pita1. Among of them, 62.5, 56.3, 58.5, 46.7, 72.4, and 57.4% of M. oryzae carry AVR-Pita1 from northeastern, southeast, western, northwest, southwestern, and central Yunnan province, respectively. The detection rate of AVR-Pita1 was, in order: southwestern > northeastern > western > central > southeastern > northwestern Yunnan province. Moreover, in total, 18 AVR-Pita1 haplotypes encoding 13 novel AVR-Pita1 variants were identified among 60 isolates. Most DNA sequence variation was found to occur in the exon region, resulting in amino acid substitution. Six virulent haplotypes of AVR-Pita1 to Pita were identified among 60 field isolates. The AVR-Pita1 has evolved to virulence from avirulent origins via base substitution. These findings demonstrate that AVR-Pita1 is under positive selection and mutations of AVR-Pita1 are responsible for defeating race-specific resistance in nature.

scholarly journals Molecular detection of ‘Candidatus Phytoplasma australasia’ and ‘Ca. P. cynodontis’ in Iraq

2017 ◽  
Vol 63 (3) ◽  
pp. 112-119
Author(s):  
Nawres Abdulelah Sadeq Alkuwaiti ◽  
Tariq Abdulsada Kareem ◽  
Layla Jabar Sabier
Keyword(s):  
Molecular Detection ◽  
Cynodon Dactylon ◽  
Sequence Data ◽  
Bermuda Grass ◽  
Nucleotide Identity ◽  
Chain Reaction ◽  
White Leaf ◽  
Pcr Products ◽  
Total Dna ◽  
Polymerase Chain

Abstract The association of phytoplasma was investigated in symptomatic tomato (Solanum lycopersicum L.), eggplant (Solanum melongen L.), mallow (Malva spp.) and Bermuda grass (Cynodon dactylon L.) plants exhibiting witches’ broom and white leaf diseases, respectively. Total DNA was extracted from tomato (n=3), eggplant (n=2), mallow (n=2) and Bermuda grass (n=8) samples. Direct polymerase chain reaction (PCR) was performed using P1/P7 primer set, then PCR products were sequenced. Sequences obtained from tomato, eggplant and mallow shared 99% maximum nucleotide identity with phytoplasma belonging to subgroup 16SrII-D, and resulted therefore ‘Candidatus Phytoplasma australasia’-related. Sequences obtained from Bermuda grass showed 100% maximum nucleotide identity to 16SrXIV-A subgroup and were ‘Ca. P. cynodontis’-related. The study presents the first molecular confirmation and sequence data of presence of ‘Ca. P. australasia’ and ‘Ca. P. cynodontis’ in Iraq.

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