Detection of pathogens from periodontal lesions

OBJECTIVE: To comparatively detect A. actinomycetemcomitans and F. nucleatum from periodontal and healthy sites. METHODS: Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed. Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR. Conventional biochemical tests were used for bacteria identification. RESULTS: A. actinomycetemcomitans and F. nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects. Among A. actinomycetemcomitans isolates, biotype II was the most prevalent. Primer pair AA was 100% sensitive in the detection of A. actinomycetemcomitans from both subject groups. Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples. Primer pair FN5047 was more sensitive to detect F. nucleatum in patients or in healthy samples than primer 5059S. Primers ASH and 5059S were more specific in the detection of A. actinomycetemcomitans and F. nucleatum, respectively, in patients and in healthy subject samples. CONCLUSIONS: PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases. The method's sensitivity and specificity is conditioned by the choice of the set of primers used.

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DETECTION AND QUANTITATION OF RED COMPLEX BACTERIA IN SUBGINGIVAL PLAQUE BY USING FLUORESCENT IN SITU HYBRIDIZATION (FISH)

International Journal of Research -GRANTHAALAYAH ◽

10.29121/granthaalayah.v5.i11.2017.2354 ◽

2017 ◽

Vol 5 (11) ◽

pp. 279-289

Author(s):

Kishore G. Bhat ◽

Aradhana Chhatre ◽

Vijay M. Kumbar ◽

Manohar S. Kugaji ◽

Sanjeevani Patil

Keyword(s):

In Situ Hybridization ◽

Diagnostic Tool ◽

Fluorescent In Situ Hybridization ◽

High Sensitivity ◽

Clinical Samples ◽

Subgingival Plaque ◽

Periodontal Pathogens ◽

Significant Difference ◽

Strong Linear Relationship

Motivation/Background: Red complex bacteria are proven periodontal pathogens. In dentistry, there is a need to identify and quantitate the organisms from the diseased sites quickly and reliably. Since culture requires several days, molecular methods are being used frequently to detect these bacteria. Among them, Fluorescent in situ hybridization (FISH) is rapid, sensitive and quantitative. An attempt is made here to evaluate the applicability of this technique as a diagnostic tool in periodontology. Method: Subgingival plaque was collected from participants, fixed with paraformaldehyde and subjected to FISH. Fluorescently labeled oligonucleotide probes were used for hybridization. After the procedure, the fluorescently stained bacteria were identified and counted from the smear and quantitated using a simple grading. Results: There was a significant difference in the prevalence and numbers of red complex bacteria in healthy and diseased subjects. A strong linear relationship existed between P. gingivalis, T. forsythia and T. denticola. Conclusions: The procedure used in the study is simple, rapid and can be easily adaptable. It also has a high sensitivity and has the ability to detect a single bacterial cell. The method can be directly applied to the clinical samples and can be used as a rapid diagnostic tool in periodontics.

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Periodontal Status and Subgingival Biofilms in Cystic Fibrosis Adults

Polish Journal of Microbiology ◽

10.33073/pjm-2019-040 ◽

2019 ◽

Vol 68 (3) ◽

pp. 377-382 ◽

Cited By ~ 3

Author(s):

TAMARA PAWLACZYK-KAMIEŃSKA ◽

RENATA ŚNIATAŁA ◽

HALINA BATURA-GABRYEL ◽

MARIA BORYSEWICZ-LEWICKA ◽

SZCZEPAN COFTA

Keyword(s):

Cystic Fibrosis ◽

Clinical Symptoms ◽

Periodontal Diseases ◽

Adult Patients ◽

Periodontal Pathogens ◽

Rt Pcr ◽

Pocket Depth ◽

Periodontal Status ◽

Periodontal Tissues ◽

Probing Pocket Depth

The aim of this study was to assess the periodontal status of cystic fibrosis (CF) adult patients and to evaluate whether there is a correlation between the bacterial population of the subgingival biofilm and the health status of the periodontal tissues in this group of adults. The study involved 22 cystic fibrosis adult patients. The periodontal condition was assessed using Plaque Index (PLI), Gingival Index (GI), and Probing Pocket Depth (PPD). The gingival sulcus samples were analyzed by the Real-Time PCR assay (RT-PCR). Majority of patients showed moderate or severe bacterial dental plaque accumulation, but none of them had clinical symptoms of periodontal diseases. RT-PCR showed the presence of periopathogens in 50% of patients. Red complex microorganisms were detected in 9.09%, orange complex in 27.27%, and green complex in 31.82% of the samples analyzed. In cystic fibrosis patients colonized by periopathogens, the periodontal markers were significantly higher in comparison to not colonized by periopathogens patients. Despite the widespread presence of bacterial dental deposits in the cystic fibrosis adult patients examined, none of them has clinical symptoms of periodontal disease; however, the presence of periodontal pathogens in subgingival biofilm may represent a possible risk factor of this disease in the future. An unsatisfactory level of oral hygiene in any patient with cystic fibrosis indicates a need to focus on standards of dental care for such patients.

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Identification Pseudomonas aeruginosa by OprD Gene for Differentiation from Other Pseudomonas Species that Isolated from Clinical Samples

International Journal of Drug Delivery Technology ◽

10.25258/ijddt.9.1.8 ◽

2019 ◽

Vol 9 (01) ◽

pp. 46-50

Author(s):

Ashwak B Al-Hashimy ◽

Huda S Alagely ◽

Akeel K Albuaji ◽

Khalid R Majeed

Keyword(s):

Pseudomonas Aeruginosa ◽

General Hospital ◽

Culture Media ◽

Final Diagnosis ◽

Clinical Samples ◽

Bacterial Isolates ◽

Molecular Method ◽

Biochemical Tests ◽

Pseudomonas Species ◽

Specific Sequences

The present study included the collection of 100 samples from various clinical sources for investigating the presence of P. aeruginosa in those sources, the samples have been collected from some hospitals in Baghdad and Hillah city (Al-qassim General Hospital, ,Al-hillah teaching hospital,and Al-hashimya General hospital ) which included wounds, burns, ear and sputum infections. The study was carried out through October 2017 till the end of March 2018. The samples were identified based on the morphological and microscopically characteristics of the colonies when they were culturing or number of culture media as well as biochemical tests, molecular identification were also used as a final diagnostic test for isolates that were positive as they belong to P.aeruginosa bacteria during previous tests based on the OprD gene which has specific sequences for P.aeruginosa bacteria as a detection gene and also consider as virulence factor so it have a synonyms mechanism to antibiotic resistance . The results of the final diagnosis showed that 38 isolates belong to target bacteria were distributed as 18 of burns, 11 isolates of wounds, 6 isolates of ear infection and 3 isolates of sputum, The examination of the sensitivity of all bacterial isolates was done for elected 38 isolation towards the 9 antibiotic by a Bauer - Kirby and the isolates were resistant for a number of antibiotics used such as Ciprofloxacin 65.7%, Norflaxacin 71%, Imipenem 63.1% Meropenem 68.4%, Gentamicin 65.7%, Amikacin 26.3%, Cefepime 68.4%, Ceftazidime 65.7% and Piperacillin 57.8%.Molecular method , All isolates (38) of P. aeruginosa positive for the diagnostic special gene (OprD) genes (100%).

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Comparison of Sampling Techniques For qPCR Quantification of Periodontal Pathogens

Revista de Chimie ◽

10.37358/rc.17.12.5993 ◽

2018 ◽

Vol 68 (12) ◽

pp. 2853-2856 ◽

Cited By ~ 3

Author(s):

Igor Jelihovschi ◽

Cristian Drochioi ◽

Aida Corina Badescu ◽

Raoul Vasile Lupusoru ◽

Alexandra Elena Munteanu ◽

...

Keyword(s):

Periodontal Disease ◽

Periodontal Diseases ◽

Pcr Analysis ◽

Treponema Denticola ◽

Subgingival Plaque ◽

Periodontal Pathogens ◽

Radiographic Evidence ◽

Gingival Bleeding ◽

As Species ◽

Qpcr Quantification

The diagnosis of periodontal disease is mainly based on use of clinical and radiographic evidence. In this study we employed a quantitative PCR analysis of Aggregatibacter actinomycetemcomitans and Treponema denticola as species strongly involved in periodontal diseases, burden in periodontal pockets to detect the main sampling factors that interfere with qPCR results. From 22 patients with advanced periodontal disease, subgingival plaque was comparatively collected by paper points and periodontal Gracey curettes. Samples were collected from the same situs in presence of gingival bleeding and absence of bleeding. The concordance and agreement of results between samples were assessed. The present study demonstrates that subgingival plaque sampling with sterile absorbable paper points is often accompanied by gingival bleeding resulting in quantification biases of periodontal pathogens.

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The SCCmec Types and Antimicrobial Resistance among Methicillin-Resistant Staphylococcus Species Isolated from Dogs with Superficial Pyoderma

Veterinary Sciences ◽

10.3390/vetsci8050085 ◽

2021 ◽

Vol 8 (5) ◽

pp. 85

Author(s):

Yuttana Chanayat ◽

Areerath Akatvipat ◽

Jeff B. Bender ◽

Veerasak Punyapornwithaya ◽

Tongkorn Meeyam ◽

...

Keyword(s):

Antimicrobial Susceptibility ◽

Sccmec Type ◽

Small Animal ◽

Control Measures ◽

Infection Prevention And Control ◽

Clinical Samples ◽

Biochemical Tests ◽

Methicillin Resistant ◽

Type V ◽

Staphylococcal Cassette Chromosome

This study characterizes clinical methicillin-resistant staphylococcal (MRS) isolates obtained from superficial pyoderma infections in dogs. Our interest was to determine the staphylococcal cassette chromosome mec (SCCmec) type and the antimicrobial susceptibility among MRS isolates from clinical cases. Skin swabs were collected and cultured. Staphylococcus species were identified and characterized with biochemical tests and MALDI-TOF-MS and antimicrobial susceptibility testing by disk diffusion. mecA detection and staphylococcal cassette chromosome mec (SCCmec) typing were achieved by PCR. Of the 65 clinical samples, 56 (86.2%) staphylococcal infections were identified. Twelve (21%) of 56 isolates were MRS infections. All MRS isolates were multidrug resistant. The ccrC and class-C2 mec, which were SCCmec type V, were the most prevalent (66.7%) among the 12 MRS isolates. The predominant SCCmec type V was found in S. aureus, S. intermedius group, S. lentus, S. xylosus, and S. arlettae. Treatment failure is a concern with the emergence of highly resistant MRS in dogs associated with superficial pyoderma. The detection of type V SCCmec MRS has previously been reported among veterinarians and dog owners but not in Northern Thailand. These infections serve as a reminder to improve infection prevention and control measures including reducing environmental contamination and potential zoonotic exposures to MRS. In addition, educational awareness of these risks in small animal hospitals needs to be increased among veterinary hospital staff, clients, and patients.

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Safety assessment and diagnostic evaluation of patients undergoing contrast-enhanced urosonography in the setting of vesicoureteral reflux confirmation

Clinical Hemorheology and Microcirculation ◽

10.3233/ch-219110 ◽

2021 ◽

pp. 1-9

Author(s):

Constantin A. Marschner ◽

Vincent Schwarze ◽

Regina Stredele ◽

Matthias F. Froelich ◽

Johannes Rübenthaler ◽

...

Keyword(s):

Urinary Tract Infection ◽

Urinary Tract ◽

Tract Infection ◽

Vesicoureteral Reflux ◽

Sensitivity And Specificity ◽

Diagnostic Tool ◽

High Sensitivity ◽

Upper Urinary Tract ◽

Clinical Diagnostics ◽

Voiding Cystourethrography

BACKGROUND: Vesicoureteral reflux (VUR) represents a common pediatric anomaly in children with an upper urinary tract infection (UTI) and is defined as a retrograde flow of urine from the bladder into the upper urinary tract. There are many diagnostic options available, including voiding cystourethrography (VCUG) and contrasted-enhanced urosonography (ceVUS). ceVUS combines a diagnostic tool with a high sensitivity and specificity which, according to previous study results, was even shown to be superior to VCUG. Nevertheless, despite the recommendation of the EFSUMB, the ceVUS has not found a widespread use in clinical diagnostics in Europe yet. MATERIALS AND METHODS: Between 2016 and 2020, 49 patients with a marked female dominance (n = 37) were included. The youngest patient had an age of 5 months, the oldest patient 60 years. The contrast agent used in ceVUS was SonoVue®, a second-generation blood-pool agent. All examinations were performed and interpreted by a single experienced radiologist (EFSUMB Level 3). RESULTS: The 49 patients included in the study showed no adverse effects. 51%of patients (n = 26) were referred with the initial diagnosis of suspected VUR, while 49%of patients (n = 23) came for follow-up examination or to rule out recurrence of VUR. The vast majority had at least one febrile urinary tract infection in their recent medical history (n = 45; 91,8%). CONCLUSION: ceVUS is an examination method with a low risk profile which represents with its high sensitivity and specificity an excellent diagnostic tool in the evaluation of vesicoureteral reflux, especially in consideration of a generally very young patient cohort.

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TO EVALUATE THE SENSITIVITY AND SPECIFICITY OF CSF-ADA AS A DIAGNOSTIC TOOL IN TUBERCULOSIS MENINGITIS

Journal of Evolution of Medical and Dental Sciences ◽

10.14260/jemds/1085 ◽

2013 ◽

Vol 2 (32) ◽

pp. 5966-5970

Author(s):

Manish Kumar Singh ◽

Chandran K ◽

Shrawan Kumar ◽

Pranjal Pankaj ◽

Ishan Parasher

Keyword(s):

Sensitivity And Specificity ◽

Diagnostic Tool ◽

Tuberculosis Meningitis

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Detection of LytA Genes in Streptococcus pneumoniae Isolated from sputum pneumonia patients

Biomedika ◽

10.31001/biomedika.v13i1.718 ◽

2020 ◽

Vol 13 (1) ◽

pp. 23-30

Author(s):

Mustika Sari Hutabarat ◽

Firdaus Hamid ◽

Irawaty Djaharuddin ◽

Alfian Zainuddin ◽

Rossana Agus ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

False Negative ◽

Pneumococcal Infection ◽

Diagnostic Methods ◽

Molecular Diagnostic ◽

Clinical Samples ◽

Biochemical Tests ◽

Negative Results ◽

False Negative Results ◽

Polymerase Chain

Streptococcus pneumoniae (pneumococcus) is a Gram-positive facultative anaerobic bacterium that is a major cause of morbidity and mortality worldwide. But the lack of reporting of disease by this bacterium in Indonesia, one of the causes is because the diagnosis of pneumococcal infection is often clinically not typical and conventional methods which are still the standard gold method often give false-negative results. So the purpose of this study was to evaluate the performance of culture and molecular diagnostic methods using the Polymerase Chain Reaction (PCR) technique in detecting Streptococcus pneumoniae in sputum clinical samples using the Autolysin (LytA) gene which is a virulence factor of this bacterium. 57 isolates from 60 samples were confirmed as Streptococcus sp through microscopic identification, culture, and biochemical tests. Then the sensitivity test with an optochin test of 9 (9%) compared the results descriptively with the PCR technique using the Autolysin A (LytA) gene which was obtained more sensitive by 15 (25%).

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Antibiotic Susceptibility Pattern of Nosocomial Isolates of Staphylococcus aureus in a Tertiary Care Hospital, Nepal

Journal of Nepal Medical Association ◽

10.31729/jnma.190 ◽

2009 ◽

Vol 48 (175) ◽

Cited By ~ 1

Author(s):

Bidya Shrestha ◽

B M Pokhrel ◽

T M Mohapatra

Keyword(s):

Staphylococcus Aureus ◽

Methicillin Resistant Staphylococcus Aureus ◽

Tertiary Care ◽

Test Method ◽

University Teaching ◽

Clinical Samples ◽

Resistance Pattern ◽

Care Hospital ◽

Biochemical Tests ◽

Methicillin Resistant

Introduction: Methicillin resistant Staphylococcus aureus (MRSA), the most common cause ofnosocomial infection has been a major cause of morbidity and mortality around the world. They arenormally resistant to most of the antibiotics used in clinical practice. This study has been carried outto fi nd out the resistance pattern among S. aureus.Methods: During November 2007 to June 2008, clinical samples from patients with nosocomialinfection were processed for culture and sensitivity following standard methodology in microbiologylaboratory, Tribhuvan University teaching hospital, Kathmandu, Nepal.Results: Among 149 Staphylococcus aureus isolates, highest resistance was observed against Penicillin(91.94%) followed by Fluoroquinolone (61.74%), Erythromycin (52.94%), Gentamicin (46.98%),Cotrimoxazole (42.95%), Tetracycline (40.94%) and others, whereas susceptibility was observedmaximum against Chloramphenicol (94.85%) followed by Rifampicin (92.61%), Tetracycline(59.06%), Cotrimoxazole (57.04%), and others. None of the isolates were resistant to Vancomycinand Teicoplanin. Of these isolates 44.96 % of the isolates were Methicillin resistant S. aureus (MRSA).Resistance to Penicillin, Fluoroquinolone, Erythromycin, Gentamicin, Co-trimoxazole and Tetracyclinewere associated signifi cantly with MRSA isolates (X2= 8.779, p<0.05, X2= 74.233, p<0.05, X2= 84.2842,p<0.05, X2= 108.2032, p<0.05, X2= 88.1512, p<0.05 and X2= 79.1876, p<0.05 respectively). Althoughmost of the Methicillin sensitive S. aureus (MSSA) isolates were susceptible to both Rifampicinand Chloramphenicol, only Rifampicin susceptibility was signifi cantly associated with them (X2=10.1299, p<0.05). Among three Biochemical tests for the detection of β lactamase detection namelychromogenic, iodometric and acidimetric test, chromogenic test method had highest sensitivity andspecifi city.Conclusions: Since MRSA comprised a greater part of S. aureus isolates and were multi-resistant,patients infected by such strains should be identifi ed and kept in isolation for hospital infectioncontrol and treated with second line of drug like vancomycin.Key Words: β lactamase, methicillin resistant Staphylococcus aureus,methicillin sensitive Staphylococcus aureus, resistance pattern

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Common ECG Lead Placement Errors. Part II: Precordial Misplacements

International Journal of Medical Students ◽

10.5195/ijms.2014.96 ◽

2014 ◽

Vol 2 (3) ◽

pp. 99-103 ◽

Cited By ~ 8

Author(s):

Allison V. Rosen ◽

Sahil Koppikar ◽

Catherine Shaw ◽

Adrian Baranchuk

Keyword(s):

Healthy Subject ◽

Patient Care ◽

Diagnostic Tool ◽

Pathological Findings ◽

Precordial Lead ◽

Single Male ◽

Lead Placement ◽

Optimal Patient Care ◽

Ecg Leads ◽

Ecg Interpretation

Background: Electrocardiography is a very useful diagnostic tool. However, errors in placement of ECG leads can create artifacts, mimic pathologies, and hinder proper ECG interpretation. This is the second of a two-part series discussing how to recognize and avoid these errors. Methods: 12-lead ECGs were recorded in a single male healthy subject in his mid 20s. Various precordial lead misplacements were compared to ECG recordings from correct lead placement. Results: Precordial misplacements caused classical changes in ECG patterns. Techniques of differentiating these ECG patterns from true pathological findings were described. Conclusion: As in Part I of this series, recognition and interpretation of common ECG placement errors is critical in providing optimal patient care.

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