When dentist harvests plaque, they insert multiple paper points into sulcus, then Clinident counts(calculate) the number of bacteria. The dentist is asking if you calculate the average value or maximum value.
The dentist insert multiple paper points to collect the maximum of “liquid” into the sulcus.
The multiple paper points are pooled and extracted all together. So the calculation represent the addition of all bacteria collected by the multiple paper point.
This mean the average of all paper points used on the same sample to be analysed.
Why Clinident can perform Real-time PCR test in 2 days, and also asking accuracy, such as comparison with other test.
About the PCR.
At the sample reception (T0 – day one) in our facility in the morning , we are performing the following step :
11 am : DNA extraction with automated platform. It take about 2 hours.
14 am : preparation of the PCR mix with an automated platform . It take about 30 minutes.
15 am : PCR reactions are performed on the PCR machine. We are performing 1 run per bacteria species ( for a maximum of 70 samples per run). Each run is about 90 minutes per machine. We have multiple machines in our facility. Several run are performed during the night.
Day 2, all run are terminated in the morning, analyzed by the software of the PCR machine and verified by the team before transfer for final control and redaction.
Report are send by email day 2 or on the morning day 3.
Is there is any word of caution before picking plaque ? Such as antibiotic, mouse wash, etc..
With paper point, the dentist will collect sulcus liquid not plaque on the teeth surface.
However, the dentist has to remove first the supra gingival plaque by mechanical action ( ex: ultra sound scaling ) and he can use at this stage bactericide to clean up the surface of the teeth and the oral cavity.
Then, he dry the zone to be analyzed and pick up the sulcus liquid with the paper point.
There is several bactericide (with chlorexidin) that could be recommended. Eludril from Pierre Fabre, Elmex has got one, Sunstar also (Perioclean)…
Antibiotic in Japan
Please refer to the below articles we are using for recommendation worldwide.
J Periodontal Res. 1995 Jul;30(4):290-3.
In vitro activity of tetracyclines, macrolides, quinolones, clindamycin and metronidazole against periodontopathic bacteria.
Miyake Y1, Tsuruda K, Okuda K, Widowati, Iwamoto Y, Suginaka H.
seems the reference use by Dr Yoda for the recommendation in Japan with tetracycline, Penicillin and New Quinolone.
We also are following recommendation provided by Prof Slots in the 2004 report.
These articles are available on the Noveo web site.
Bacteria account: There is explanation about “E= Exponential ex) 1.2E+107 =12000000 “ in the Clinical Report. What is the calculation of the bacterias :
Exponential is a mathematical way to express numbers in a short way.
for example :
- 1E+01 = 1E+101 = 10
- 1E+02 = 1E+ 102 = 100
- 2E+02 = 2E +103 = 2000
- 2,3E+03 = 2.3E+ 103 = 2300
- 1,2E+07= 1.2E+107 =12 000 000
Bacteria account: In the table for Microbiological results, the total number of pathogenic organism is not equal to total bacteria count.
There is more than 200 different bacterial species into oral cavity (not only periodontal pathogenic Aa, pg…) but also healthy and commensal bacteria.
So we are also measuring the total bacteria count for all bacteria present into the sample.
Total bacteria count do not represent the addition of the pathogenic bacteria but addition of all bacteria present into the sample (including good bacteria necessary for microbial equilibrium).
The ratio (percentage) between pathogenic bacteria and total bacteria count give information about the “competition” situation into the periodontal pocket between pathogenic and other bacteria.
A high percentage indicate that the pocket is highly contaminated by a specific pathogenic bacteria strain and that competition with “good & healthy bacteria “ is limited.
DNA amplification: How many DNA amplification cycles ?
DNA amplification is done by real time PCR (Polymerase Chain Reaction).
Please find a link to wiki and a specific tutorial :
- https://en.wikipedia.org/wiki/Real-time_polymerase_chain_reaction
- http://www.microbiologybook.org/pcr/Real%20Time%20PCR%20Tutorial.pdf
We are performing 40 DNA amplification cycle during real time PCR . It take about 90 minutes for the all process.
The full processing (DNA extraction + DNA amplification + final analysis take about 5 to 6 hours.
Please refer to the Pr Luigi Canullo, this article is available on the Noveo web site :
“ Microbiological assessment of the implant-abutment interface in different connections: cross-sectional study after 5 years of functional loading “
Threshold: Information about the pathogen loan threshold
If the therapist is able to rapidly decrease and maintain at low levels the counts of periodontal pathogens, this will lead to a reduction in the counts of the entire ecosystem and improvement in clinical parameters.
The Perio-Analyse test examines the situation and the changes that occur in the sub gingival microbiota resulting from periodontal disease and periodontal treatment to determine the sequence of changes that takes place in the sub gingival microbiota that lead to periodontal stability during treatment.
Treatment is mechanical treatment and surgery that could be associated with specific systemic antibiotic.
Mechanical therapies, including scaling and root planing and surgery associated with bactericide will reduce the microbial load.
Antibiotic should be use with caution in the molecule selection to be efficient and to avoid bacteria resistance.
The following articles are available on the Noveo web site :
Haffajee A et al (2000) described that red complex bacteria group (Pg, Tf , Td) , under certain value antibiotic treatment seems not really impact the mean pocket depth reduction compared to surgery.
The pathogen load threshold proposed in Perio-Analyse is adapted from Teles RP et al (2000) review.
Haffajee AD, Teles RP, Socransky SS.
The effect of periodontal therapy on the composition of the sub gingival microbiota.
Periodontal 2000. 2006;42:219-58
Teles RP, Haffajee AD, Socransky SS.
Microbiological goals of periodontal therapy.
Periodontal 2000. 2006;42:180-218
Do you have any sample article for the paper to explain the method of analyzing the bacteria by Perio-Analyse. She would like to know how to write the method of Perio-Analyze in paper.
Ex. “extract the DNA and…”, “duplicate, triplicate… ” and “centrifuge them XX minutes”
Canullo L, Penarrocha-Oltra D, Soldini C, Mazzocco F, Penarrocha M, Covani U.
Clin Oral Implants Res. 2015 Apr;26(4):426-34.
How long you will store the samples after analysis?
We store DNA sample for 5 years
Do you know any study used Perio-Analyse that shows the effect of chronologic and temperature changes to DNA samples?
There is studies about paper point evaluation (Oral Microbial Immunol. 1999 Oct;14(5):326-30. Sampling of periodontal pathogens by paper points: evaluation of basic parameters.Hartroth B1, Seyfahrt I, Conrads G.).
There is no published studies about the chronologic stability of the DNA on paper point at difference temperature. However, we performed (for the CE mark registration) internal validation about the stability of DNA on paper point during several weeks. We found a good stability during more than 1 month. However, we decided to recommend to not exceed 2 weeks shipment at room Temperature.
Why not only collecting the saliva ?
Because the presence of periodontal bacteria into the saliva is a risk factor, but not a diagnostic. Only presence of non equilibrated microflora in the sulcus could be considered as a diagnostic tool and should be treated.
How can I use Perio-Analyse in my practice ?
- For new patient
- Patient with periodontal disease to be treated
- Patient at risk, going to have dental implant after periodontal disease
- Patient at risk having dental implant (annual control)
- Patient with dental implant having mucositis or peri implantitis
- Patient at genetic risk (Interleukin mutation)
- Diabetic patient
- Pregnant patient having periodontal disease risk
Why Ab is associated with surgery/mechanical treatment ?
Aa is intracellular bacteria. Certain bacteria like AA and Pg are not strict anaerobic and could survive in presence of oxygen after surgery
Why Candida albicans could be at risk ?
Because in presence of antibiotic and after bacteria reduction and limited competition , Ca can grow and fully colonize the sulcus.
What are the unit use for bacteria load ?
10 000 = 10 4 = E+04 = 10 000 bacteria in the tested sulcus
What is PCR ?
- PCR is amplification of specific zone of the DNA
- Each amplified zone is specific from a bacteria species
How long is the PCR process ?
- DNA extraction 2 hours
- PCR preparation 1 hour
- PCR run 2 hour (about 10 patients per run)
When should I test for the second time ?
- 3 months after the first sampling and antibiotic use
Antibiotic recommendation:
- Aa alone or with red complex < the threshold = New Quinolone
- Aa + red complex > the threshold = Penicillin
- Absence of Aa other bacteria > the threshold = Tetracycline
How many paper point to be used ?
Minimum 2 per site. 10 paper points for a pool
How many samples can by run per day by Clinident ?
about 150/day
Vacation/Holiday schedule of Clinident ?
Lab is is closed 2 weeks per year (1 week in August and 1 week end of December)
What are the DNA target and DNA standard of the PCR developed by Clinident ?
- Ribosomal 16S DNA sequences
- Calibrated DNA from DSMZ (Germany) and Institut Pasteur (France)
About Unit Oral
Why should I check the water unit?
The water in your practice is characterized as “process” and therefore must comply with the Health and Safety Legislation. Part of this legislation is regular monitoring of the microbiological quality.
Is there new EC or national directive for prevention in oral health care practices demand?
Yes after Germany, a new directive was published in early April 2016 in the Netherlands. The regulations regarding water quality is further explored in this new directive. One of the changes is that the minimum water quality should be checked every six months
What is the best time for sampling?
The best time for sampling is in the morning before entering the first patient. After a (chemical) disinfection wait for at least 48 hours before the water unit can be tested.
How often should I perform a DUW test?
Every month is the best option to secure the dental practice. Every 6 months is the existing directive
Is it possible to remind me that it's time for a sampling?
Yes, If you will need this, please let us know .
All units must be sampled?
Yes, all units in the practice should be sampled.
I have a separate bottle system, do I have to sample?
Also, with a bottle system, there is a risk of contamination, for example during the filling of bottles or by recoil of micro-organisms from the mouth of a patient. Bottle systems should be monitored for the quality of the water unit.
How soon will I get the results?
Within a week after receipt of the collected water in the laboratory, we will send you the results.
I receive the results by e-mail?
The result will be sent by e-mail and addressed to the contact person indicated on the order form. The result we will never communicate to third parties without explicit consent of the client.
Do you have any literature studies on water quality in dental practice?
About Perio-Analyse
If you need more background information, please contact us. We will send you more information or literature.