Study On The Application Of Fungal Fluorescence Staining In The Diagnosis Of Sporotrichosis

Background:Sporotrichosis is a chronic mycosis caused by pathogenic schenkerosporium(pathogenic Sporothrix schenckii),which mainly invades the skin,subcutaneous tissue and nearby lymphatic vessels.The common diagnostic methods of the disease are fungal culture and histopathological examination.These two methods take a long time,complex operation and high technical requirements.and more rapid and simple detection methods are needed in clinic.Fungal fluorescence staining is a kind of fluorescent staining method based on calcium fluorescent white reagent,which can stain the cell wall of fungi quickly and specifically.This method was initially mainly used for direct microscopic examination of superficial fungal infections.Recent studies have shown that it can be used for staining deep skin lesions and paraffin sections to achieve rapid and specific detection of pathogenic fungi.However,the application of this method in sporotrichosis is less reported.Objection:1.Compare the positive rate of fungal fluorescence staining and superiodate-schiff(PAS)staining in tissue sections of sporotrichosis,and explore the application of fungal fluorescence staining in pathological diagnosis;2.To analyze the relationship between the number of yeast cells and clinical classification in the pathological sections of sporotrichosis,and to explore the factors affecting the clinical manifestations of sporotrichosis;3.Fungal fluorescence staining solution was used to detect the skin lesions of patients with sporotrichosis,to observe the detection of yeast cells,and to explore the application of fluorescence staining in the diagnosis of sporotrichosis.Method:1.Paraffin sections of pathological tissue were selected: review of the diagnosis of sporotrichosis in our department from November 2019 to December 2020.The inclusion criteria meet one of the following criteria:Fungi culture positive and identified as sporotrichosis;The fungal culture was negative,the clinical manifestations were in accordance with the typical skin lesions of each clinical type of sporotrichosis,the histopathology was in accordance with the tissue image of deep fungal infection,and the systemic antifungal therapy was effective.2.Fungal fluorescence staining and PAS staining were compared in pathological sections of sporotrichosis by detecting yeast cells;paraffin sections of fixed type and lymphatic type were selected in fungal culture positive cases of sporotrichosis.After fluorescence staining,the amount of yeast cells in every 10 fields of vision was observed and recorded.3.This project is approved by the Ethics Committee of the first Hospital of Jilin University.4.Statistical method: the composition of counting data was compared and applied χ2test,and the rank data was applied rank sum test.The p<0.05 was statistically significant.Results:In this study,136 cases of standard cases were collected,including 123 cases of sporotrichosis,8 cases of fresh tissue of sporotrichosis and 5 cases of non-fungal infection with other skin diseases.1.Among 123 cases diagnosed as sporotrichosis,105 cases were positive for fungal culture(positive rate 85.37%),87 cases were positive for fungal fluorescence staining in paraffin sections(positive rate 70.73%),and 50 cases were positive for PAS staining(positive rate 40.65%).Of 105 positive fungal cultures,77(73.33%)were positive for fluorescence staining,and 48(45.71%)PAS positive staining were calculated by paired chi-square,the difference was statistically significant.2.Of 105 cases of fungal culture positive and identified as sporotrichum,77 cases(positive rate 73.33%)and 48 cases(positive rate 45.71%)were positive for PAS staining in paraffin sections.The data analysis showed p<0.05,the difference was statistically significant.3.In 18 cases of fungal culture negative but with diagnostic criteria for sporotrichosis,Fungal fluorescence staining in paraffin sections was positive in 10 cases(positive rate 55.56%),Negative staining in 8 cases(negative rate 44.44%);PAS staining positive 2 cases(positive rate 11.11%),Negative 16 cases(negative rate 88.89%).Data analysis showed p>0.05,The difference was not statistically significant.4.In fungal-positive paraffin sections,38 and 55 fixed types were selected,including paraffin sections < 5 / 10 horizons,26,yeast cells> 10 / 10 horizons,9,5-10 / 10,3,yeast cells < 5 / 10,44,yeast cells> 10 / 10,5,5-10 / 10,6.Data analysis found that although the number of yeast cells in fixed tissue paraffin sections was less than lymphocytes,the difference was no significant(p> 0.05).5.The results showed that the detection rate of yeast cells was 100% in the skin lesions of patients with sporotrichosis and other skin diseases,but no yeast cells were found in other skin diseases.Conclusions:1.Fungal fluorescence staining was better than PAS staining in paraffin sections of pathological tissues of sporotrichosis patients,suggesting that fungal fluorescence staining could be used for pathological tissue staining of sporotrichosis as an auxiliary diagnosis of sporotrichosis.2.There was no significant difference in the number of yeast cells in paraffin sections of different clinical types.It was inferred that the clinical manifestations of sporotrichosis were independent of their number.3.In the pus of patients with sporotrichosis,the positive rate of yeast cells detected by fungal fluorescence staining microscopy is high.In clinical diagnosis,fluorescence staining can be considered for direct microscopic examination.