Handbook For Microbiology Practice In Oral And Maxillofacial Diagnosis
A study guide to laboratory techniques in Oral Microbiology
by
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1. Disc diffusion method:
For determing the drug susceptibility of the isolate there are two techniques.
a. The stoke’s method:
- Although in the past this method was the technique of choice but as this method has some limitations, so at present it has only a historical importance.
b. Modified Kirby – bauer method:
- On world wide scale the most popular method is the Kirby bauer method. It was subsequently modified and updated by the NCCLS ( National Committee for Clinical Laboratory Standards now named as CLSI, USA.)
- The zone diameters that are generated by the test are meaningless without reference to the minimum inhibitory concentration ( MIC ) correlates and interpretative guidelines, published by the CLSI. ( see Table NCCLS )
Preparation of culture plates:
1. Preparation of plates.
2. Preparation of inoculums.
3. Inoculation of inoculums
4. Incubation
5. Interpretation
6. Results.
1. Preparation of plates:
- Medium allowed to cool to 50°c and add 5% blood agar.
- Pour the medium in petri dishes to a depth of 4mm ( 25ml in a 9cm dish.)
- Before inoculation dry the plate with lid agar so that there are no droplets of moisture on the agar surface.
2. Preparation of inoculums:
- Take 2-3 similar type of colonies and incubate in nutrient broth an incubate 3-4 hrs at 35-37°c.
- Transfer a loopful of control inoculums to 1/3 sides of pre dried medium plate.
- Antibiotic discs are applied with forceps on the junctional line between the test and control organisms.
- 4 disc on 9cm dish / 6 discs in 10cm dish.
3. Inoculation of inoculums:
- Inoculum from the culture plates are transferred to plate by loops.
- Well formed colonies are taken, & test and control both can be added in a same plate.
4. Incubation:
- Test & control is allowed are incubated overnight at 35°c - 37°c.
5. nterpretation of plates:
- Interpretation is based on zone of inhibition around the disc and maximum zone in mm is noted.
- Results are given as either “resistant” or “sensitive” strains of organisms.
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About the Book
Oral Microbiology is a study of microbial diseases of the oral cavity. For the depth and precision of knowledge in this noble field, it can be divided into clinical and practical aspects of microbiology. Oral microbiology denotes the congregation of basic medical sciences and practicing dentistry. The most common oral microbial disease and present ever since the olden days of earth has been documented about dental caries. However, there are many other microbiological diseases that affect the oral cavity. These microbial diseases can cause potential tissue damage; or the majority of the time it leads to compromised oral health; also sometimes it can escort to death. The state of morbidity and mortality factors associated with these microbial diseases leaves an important and special enlightenment of oral microbiology in terms of diagnostic procedures are needed. Since it has no technique of its own, comprehension of this special field has to be drawn and adapted from the disciplines of medical microbiology. The context of bringing this book is an attempt to get an attention towards diagnostic procedure and laboratory techniques that are emphasized over the oral microbiological practice.
About the Author
Dr. Arvind Babu Rajendra Santosh is an Indian national who pursued his undergraduate education at the Rajiv Gandhi University of Health Sciences, Karnataka, India from where he graduated with a degree in Bachelor of Dental Surgery, which he completed in 2007. Dr. Babu was employed by the Vydehi Institute of Dental Sciences and Research Center in the department of Operative dentistry and Endodontics for one academic year. He then entered the Dr. N.T.R. University of Health Sciences, Andhra Pradesh, India to pursue his post graduation in Master of Dental Surgery degree in the subject Oral and Maxillofacial Pathology and Microbiology which he completed in 2012. Dr. Babu was employed to the C.K.S. Theja Institute of Dental Sciences and Research in the department of Oral and Maxillofacial Pathology and Microbiology. He then joined the Faculty of Medical Sciences – Dental Programme, The University of the West Indies – Mona on February 1, 2013. He has been taken duties of teaching and research co-ordinator. His teaching area includes Oral pathology and Microbiology, Forensic Dentistry, Oral Histology and Oral embryology. His particular interest in dentistry lies in research in Oral cancer, Auto immune disease, Oral Microbiology and Forensic dentistry. He published papers in peer reviewed indexed journals . He has presented scientific paper and posters in dental and specialty conferences. He is a member of the International Association of Dental Research, International Association of Oral Pathology, Indian Association of Oral and Maxillofacial Pathologists, Indian Association of Forensic Odontologists and Indian Dental Association.