# Prevalence of _Trichomonas tenax_ in the Population Affected by Periodantal Disease--A Review
## 4 Discussion
Microscopic examination was the primary detection method in forty studies, with thirty-nine using light microscopy and one employing phase-contrast microscopy. The advantage of microscopy is the easy and quick visualization of trichomonads. Cultivation allows for the isolation and identification of _Trichomonas tenax_. We were able to find fourteen reports that detect the unicellular organism through cultivation in an axenic culture medium known as Diamond's medium, which was established in the second half of the 20 th century [92]. However, we acknowledge the limitations of both of these methods, namely limited specificity and low sensitivity. Cultivation is also time-consuming. There is a high probability that the results may not be completely accurate in studies that have used these two methods because of their shortcomings. Furthermore, it is possible that the samples may have been contaminated with _Trichomonas hominis_ or _Trichomonas vaginalis_, whose morphologies are particularly similar, as discussed in Table 1, causing false-positive results. As discussed earlier, most studies have examined _Trichomonas tenax_ and _Entamoeba gingivalis_ together. Diagnosis may be complicated by the tendency of _Trichomonas tenax_ to form amoeba-like forms that may be overlooked by light microscopy [81]. These forms are visually similar to those of _Entamoeba gingivalis_ and may lead to the misinterpretation of results. It should be noted that in some studies light microscopy and cultivation are complemented by Polymerase chain reaction (PCR), which increases diagnostic accuracy and reliability. PCR can detect the DNA of microorganisms even at very low concentrations, which is difficult or impossible with light microscopy. Molecular techniques are the only method used in some studies. These techniques have a high sensitivity and specificity and can be used for strain discrimination. The high sensitivity of PCR is highlighted in some studies, where it was able to detect the nucleic acid of the protozoan in samples that had previously been found to be negative by light microscopy [48,81]. The shortcomings of molecular techniques include the need for advanced laboratory skills and specialized equipment, as well as the possibility of contamination and false-positive results. We established that PCR was used in 15 studies. Some of the _Trichomonas tenax_ genes, such as the beta-tubulin gene, 18S rRNA gene, rbb1 gene, have been analyzed by PCR [42,74,93]. Loop-mediated isothermal amplification (LAMP) is a relatively new method that requires a shorter run time and is more sensitive than PCR [94,95]. There was only one study in which LAMP was employed to detect _Trichomonas tenax_, but this used canine oral samples and the specific detection of ITS (internal transcribed spacers) and the 5.8S rRNA gene [96]. We observed that gaps currently exist in immunological diagnosis. Advances in diagnostic techniques and tools may contribute to increasing the number of epidemiological studies on _Trichomonas tenax_. Despite these findings, there is still insufficient information regarding the distribution of _Trichomonas tenax_ and there is a need for future research on its prevalence. Conducting more epidemiological studies could prove to be useful in understanding its prevalence as well as the social disparities among affected populations. Furthermore, future research may focus on the pathogenesis and impact of this microorganism on oral health, its genetics and biology and its relationship with other microorganisms in the oral cavity. The development of more sensitive and specific methods for the diagnosis of _Trichomonas tenax_ may facilitate a more accurate determination of its presence in the human mouth. Improved methods could help not only in the diagnosis of existing disease, but also in the screening of people with dental calculus. Because of its advantages, we may recommend the LAMP method as an efficient screening method in the future.