Mycology Of Superficial Cutaneous, Hair And Nail Fungal Infection Among Patients Attending Dermatology Clinic At Hospital Melaka
Mycology Of Superficial Cutaneous, Hair And Nail Fungal Infection
General Information
Study Information
Dermatology
Certain skin disorders attributable to infection or infestation
Certain skin disorders attributable to fungal infection
The clinical samples that will be collected for this study will include skin scrapings, nail clippings and hair pluckings. The clinical samples will be collected after obtaining proper informed consent. The suspected dermatomycoses lesion will be cleaned with 70% alcohol. The skin scrapings will be collected from the active edges of the lesion with a sterile blunt scalpel. The infected nail will be clipped with nail clippers and infected hair will be epilated from the scalp lesion with sterile forceps. A part of each of these samples will be used for KOH mount examination and fungal culture. All samples of skin scrapping, nail clipping and hair plucking will be subjected to direct microscopic examination with 10%-20% potassium hydroxide (KOH) solution and fungal culture in Sabouraud dextrose agar tubes with chloramphenicol. For fungal culture test, the clinical samples will be seeded on the surface of agar media, incubated at 30 degree celcius and culture growth will be analyzed daily. The isolates will be identified by examination of the cultural characteristics and microscopic morphology. Cultures without growth during a period of up to four weeks were considered negative.
The remaining samples will be used for fungal DNA extraction and PCR. For fungal PCR test, conventional PCR will be used. Fungal PCR will be performed using primers that amplify the Internal Transcribed Spacer regions, (ITS1 and ITS2 regions) of the ribosomal DNA of all fungi. Internal Transcribed Spacer (ITS) is a region that are recommended for highly variable sequences of great importance in distinguishing fungal species by PCR analysis. These primers will amplify a wide range of fungal targets and work well to analyze DNA isolated from individual organisms. The ITS primers that will be used in this experiment are ITS 1 for Forward sequence of 5’-TCC GTA GGT GAA CCT GCG G-3’ and ITS 4- 5’/5Biosg/TCC TCC GCT TAT TGA TAT GC- 3’ as well as DNA amplification methods suggest in One Taq DNA Polymerase kit for 35 cycles. After PCR, the amplified DNA fragments will be detected using 1.5% agarose gels and will be exposed to UV light and photographed. The amplified DNA will be sequenced and further analyze to identify the fungal species. Results will be present in table. All excess/leftover biological samples will be discarded.
Inclusion & Exclusion Criteria
Study Timeline
Gender : Involve All Gender
Sites
HOSPITAL MELAKA
Team Members
HOSPITAL MELAKA