RAPID IDENTIFICATION OF DERMATOPHYTES IN SKIN SPECIMENS OF DERMATOPHYTIC PATIENTS BY PCR: OBSTACLES AND SOLUTIONS

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MIRZAHOSSEINI H.; BAYAT F.; RAZAGHI ABYANEH M.

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Journal: Iranian Journal of Infectious Diseases and Tropical Medicine Year:2007 | Volume:12 | Issue:36 Page(s): 39-44

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Title

RAPID IDENTIFICATION OF DERMATOPHYTES IN SKIN SPECIMENS OF DERMATOPHYTIC PATIENTS BY PCR: OBSTACLES AND SOLUTIONS

Author(s)

MIRZAHOSSEINI H. | BAYAT F. | RAZAGHI ABYANEH M. | Issue Writer Certificate

Keywords

DERMATOPHYTEQ3

PCRQ3

SKIN SPECIMENSQ3

DNA PURIFICATIONQ3

Abstract

Background and objective: Today, it is important to affect a rapid and accurate etiological identification because development of treatment strategies that target specific species of DERMATOPHYTEs. Many countries benefit from a number of different molecular techniques to routinely diagnose a wide range of infective diseases. Likewise in our country some of the researchers, through employment of some of these techniques in addition to the use of colonies from patient's skin culture, have been able to distinguish DERMATOPHYTEs sporadically. However, in this research for the first time in Iran we had the opportunity to evaluate the quick and direct specification of these fungi by using PCR on patient's skin specimen and young colony culture.This method has been tested world wide on a small scale.Material & Methods: In our present study we investigated different methods for homogenizing patient's skin specimen and deconstructing fungus cell wall along with improving the conditions for DNA PURIFICATION via common methods and available kits. In addition, in order to apply PCR, different parameters like using multiple primer pairs, DNA polymerase enzymes, changes in PCR temperatures and times program were evaluated.Results: Along the way we were faced with different obstacles such as: fungus's hard cell wall, low amounts of fungus DNA in relation to the total purified DNA, different influential factors in the DNA PURIFICATION and PCR process, for which different solutions were examined. Finally through improvement of purification conditions and considerable amounts of fungus DNA extracts from a three sample mixture of patients skin specimen (DERMATOPHYTEs the same species), a positive PCR result were found. However, this method is not a justifiable and applicable way of routinely identification DERMATOPHYTEs.Conclusions: Despite the applied strategies in this study to instill the use of PCR for quick specification of DERMATOPHYTEs in patient's skin sample and the positive results that were obtained, it seems that with respect to the limitation on the amounts of patients skin specimen and the presence of potential inhibitors of DNA polymerase action in the skin specimen, it would be better to use DERMATOPHYTE colonies in the early stages of culturing instead of the skin specimen in addition to the complementary investigations and improvement of the methods.

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APA: Copy

MIRZAHOSSEINI, H., BAYAT, F., & RAZAGHI ABYANEH, M.. (2007). RAPID IDENTIFICATION OF DERMATOPHYTES IN SKIN SPECIMENS OF DERMATOPHYTIC PATIENTS BY PCR: OBSTACLES AND SOLUTIONS. IRANIAN JOURNAL OF INFECTIOUS DISEASES AND TROPICAL MEDICINE, 12(36), 39-44. SID. https://sid.ir/paper/52955/en

Vancouver: Copy

MIRZAHOSSEINI H., BAYAT F., RAZAGHI ABYANEH M.. RAPID IDENTIFICATION OF DERMATOPHYTES IN SKIN SPECIMENS OF DERMATOPHYTIC PATIENTS BY PCR: OBSTACLES AND SOLUTIONS. IRANIAN JOURNAL OF INFECTIOUS DISEASES AND TROPICAL MEDICINE[Internet]. 2007;12(36):39-44. Available from: https://sid.ir/paper/52955/en

IEEE: Copy

H. MIRZAHOSSEINI, F. BAYAT, and M. RAZAGHI ABYANEH, “RAPID IDENTIFICATION OF DERMATOPHYTES IN SKIN SPECIMENS OF DERMATOPHYTIC PATIENTS BY PCR: OBSTACLES AND SOLUTIONS,” IRANIAN JOURNAL OF INFECTIOUS DISEASES AND TROPICAL MEDICINE, vol. 12, no. 36, pp. 39–44, 2007, [Online]. Available: https://sid.ir/paper/52955/en

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