candida culture and strain identification in patients with pss

pSS 相关口腔念珠菌的诊断和菌株鉴定

Candidiasis is caused by Candida spp., which are opportunistic pathogens. Common strains include Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, and Candida dubliniensis1. Opportunistic infections caused by Candida spp. include superficial Candida infections and invasive candidiasis. Invasive candidiasis mainly happens in immunocompromised individuals; for example, in patients with acquired immunodeficiency syndrome (AIDS), invasive candidiasis can significantly threaten the quality of life or even the life span2. Superficial Candida infection, such as mucocutaneous candidiasis, is more common in patients3. 
Oral candidiasis, a superficial Candida infection, is the most common human fungal infection. Candida albicans is a normal commensal of the mouth; the carriage rate of Candida spp. has been reported to range from 20% to 75% in the general population without any symptoms4. Overgrowth of Candida can lead to local discomfort in patients, such as the altered taste of sensation, burning sensation in the mouth, dysphagia due to poor nutrition, delayed recovery, and prolonged hospital stay. Long-term oral candidiasis can lead to severe invasive candidiasis, resulting in significant morbidity and mortality5. Oral candidiasis can also increase the risk of oral cancer5. Impaired salivary gland function is among the risk factors for oral candidiasis6. The initiation of infection is the adhesion of Candida to epithelial cell walls. This is followed by the proliferation and filamentation of Candida and the formation and maturation of biofilm7. Biofilms are extremely difficult to eradicate and are resistant to conventional antifungal treatment, making clinical treatment a challenge for biofilm-associated oral Candida infection7.
Primary Sj&#246;gren's syndrome (pSS) is an autoimmune disease that is characterized by dry mouth, dry eyes, and other systematic symptoms8. Dry mouth is the most frequent symptom of pSS. Saliva has important physiological antifungal functions. On the one hand, it can dilute and scour the mouth and thereafter remove organisms from the mucosa. On the other hand, saliva contains antimicrobial proteins, such as lactoferrin, sialoperoxidase, lysozyme, histidine-rich polypeptides, and specific anti-candidal antibodies, which can interact with the oral mucosa and prevent the overgrowth of Candida6. The reduced saliva flow may predispose pSS patients to oral candidiasis. An observational cross-sectional study conducted in 61 pSS patients found that 13.1% of pSS patients presented oral signs of candidiasis and colony forming unit (CFU)/mL of Candida albicans was found significantly and negatively correlated with levels of unstimulated whole saliva (UWS) and stimulated whole saliva (SWS)9.
The methods introduced in this manuscript can identify oral Candida infection based on characterized morphological attributes (e.g., the hyphae and yeast cells) in direct smear or after culturing10. The strain identification of the Candida using a Candida diagnostic agar is based on the formation of different colored colonies with varied morphology, which result from the cleavage of chromogenic substrates by species-specific enzymes11. In this manuscript, systematic methods for detecting oral Candida infection are introduced, including traditional oral Candida culture and rapid examination by potassium hydroxide (KOH) smear test and calcofluor white (CFW) staining assay10. Additionally, the antifungal susceptibility test, which can guide clinical oral Candida infection treatment, has been introduced. In actual situations, it is not necessary to perform all of the introduced methods in this manuscript; the oral Candida detection method(s) can be selected according to the purpose. The examination of oral Candida infection and the determination of Candida strains in pSS patients not only benefit the disease treatment but also help assess the characterization of oral candidiasis and the species profiles.

作者聚焦：口腔 念珠菌 诊断推进 pSS 患者的临床治疗方案

原发性干燥综合征患者口腔 念珠菌 感染的检查

In this research, we present a protocol for the examination of oral candida infection in patients with primary Sjogren syndrome, which can be used for timely treatment to avoid infection-related complications. We provide a series of systematic, simple, and feasible methods for detecting oral candida infections, identifying candida strains, and testing the antifungal susceptibility of commonly-used drugs in patients with primary Sjogren's syndrome. Maybe our focus on the role of oral candida infection in the onset and the progression of primary Sjogren's syndrome pathologists in our future studies.

假丝酵母属 pSS 患者的培养和菌株鉴定

首先，在 PBS 中获取患者的唾液样本。将 7 克 SD 琼脂粉溶解在 100 毫升双蒸水中。使用高压灭菌器，在 121 摄氏度下对培养基灭菌 15 分钟。将约 10 毫升培养基倒入 10 厘米的微生物培养皿中，让其冷却，然后在琼脂表面划出 200 微升的唾液样品，并在 37 摄氏度下孵育 48 小时。为了制备念珠菌诊断琼脂培养基，将4.77克粉末与100毫升双蒸水混合。将混合物加热到 100 摄氏度，直到达到轻微沸腾，然后将大约 10 毫升的培养基分配到每个 10 厘米的微生物培养皿中，并让它凝固。接下来，将念珠菌菌落转移到 500 微升 SD 培养基中，并使用移液管将它们彻底悬浮。将 50 微升悬浮液铺在诊断琼脂上，并在 37 摄氏度下将平板孵育 48 小时。最后，使用特定菌落的颜色和图案从成熟菌落中鉴定菌株。使用念珠菌诊断琼脂的念珠菌菌株鉴定试验显示，原发性干燥综合征患者有白色念珠菌、光滑念珠菌和白色念珠菌加克鲁塞念珠菌的口腔感染。

Research

JoVE Journal

Medicine

Candida Culture and Strain Identification in Patients with pSS

Examination of Oral Candida Infection in Primary Sj&#246;gren&#39;s Syndrome Patients

Primary Sj&#246;gren's syndrome (pSS) is an autoimmune disease characterized by symptoms such as dry mouth, dry eyes, and other systematic symptoms. Due ...

科研

医学

Sample Collection for Diagnosis of Oral Candida in pSS Patients

pSS 患者口腔 念珠菌 诊断的样本采集

Rapid Detection of Oral Candida Infection in pSS Patients Using KOH Smear Test and CFW Staining Assay

使用 KOH 涂片试验和 CFW 染色测定法快速检测 pSS 患者口腔 念珠菌 感染