# Molecular Techniques and Target Selection for the Identification of _Candida_ spp. in Oral Samples
## 5 Conclusions  
The oral mycobiome is intricate, dynamic and involves extensive biofilm formation. _Candida_ is frequently found in the human mouth and, as with several other pathogenic fungi, appears to be an oral resident in some individuals. Because of the similarity between species, the correct identification is difficult but crucial to the success of the therapy outcome. When choosing a technique for _Candida_ identification in clinic settings, material costs, the use of trained professionals, the complexity of the technique, the specificity of the results, and time should be taken into consideration. We focused on a broad range of molecular techniques that have been used for the identification of _Candida_ species in oral samples, having in mind that a timely and accurate diagnosis of _Candida_ infection is indispensable for timely intervention with appropriate antifungal therapy. To overcome this challenge, a fast, reliable, inexpensive, and uncomplicated point-of-care diagnosis is needed. For _Candida_ spp. identification, only a few techniques fit this criterion: from the necessary sample treatment to time-to-result, only LAMP and Multiplex PCR seem to look promising. LAMP is considered by many authors to be a highly useful diagnostic technique, especially in areas where access to complex healthcare facilities is limited. However, the amount of data and results on the efficiency of this technique when applied to the diagnosis of _Candida_ infections are still scarce when compared to other techniques. Furthermore, the clinical application of LAMP on a larger scale has yet to be achieved. LAMP requires the least amount of time out of all the techniques to reach a diagnosis and does not require costly equipment. Using the _ITS_ sequence as a target, it is possible to design primers to identify _Candida_ spp. sampled from the area of infection. Taken together, LAMP specificities and requirements seem to be the most adequate for the simplest and most time-efficient diagnostic of oral candidiasis. It is worth highlighting that the exponential growth of gene sequence databases has provided the ideal conditions to develop more efficient and reliable primer designs, enhancing target specificity and the accuracy of diagnosis. Ultimately, the pros and cons of each molecular technique detailed in this review can hopefully help dentists who deal with patients with inflammatory conditions to choose the most appropriate diagnostic method. Nevertheless, novel developments within this field of research may lead to improvements in currently available techniques and to the development of new ones. The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/app12189204/s1](https://www.mdpi.com/article/10.3390/app12189204/s1)]([https://www.mdpi.com/article/10.3390/app12189204/s1](https://www.mdpi.com/article/10.3390/app12189204/s1)), Table S1: Primers (Universal and species-specific) used for _Candida_ species detection and size of fragments separated by agarose gel electrophoresis; Table S2: List of primers used for Nested PCR; Table S3: Restriction Fragment Length Polymorphism (RFLP) fragment patterns in oral _Candida_ spp. isolates with restriction enzymes; Table S4: Oligonucleotide probes used for _Candida_ spp. in non-oral isolates; Table S5: Primers used in Kasahara (2014) for the detection of _Candida_ spp. with LAMP. **Author Contributions:** Conceptualization, A.S.D., M.J.C., and A.A.; methodology, J.M., A.S.D. and R.M.S. ; validation, J.M., M.J.C., R.M.S., A.C.E., A.A. and A.S.D. ; formal analysis, J.M. ; investigation, J.M., R.M.S. and A.S.D. ; resources, A.S.D. ; data curation, R.M.S., A.C.E., A.A. and A.S.D. ; writing--original draft preparation, J.M. ; writing--review and editing, J.M., M.J.C., R.M.S., A.C.E., A.A. and A.S.D. ; supervision, A.S.D. and A.A.; project administration, A.S.D. All authors have read and agreed to the published version of the manuscript.