激光共焦显微镜在真菌性角膜炎诊断中的应用

目的:探讨激光共焦显微镜HRT3-RCM在真菌性角膜炎诊断中的应用价值。方法:对临床拟诊真菌性角膜炎患者32例32眼行HRT3-RCM检查,同时进行角膜刮片细胞学检查。结果:在32例患者中,24例在角膜组织刮片中查到了菌丝,阳性率为75%;30例在激光共焦显微镜下观察到真菌菌丝,阳性率为94%。结论:HRT3-RCM是利用激光作为光源的新一代共焦显微镜,是一种快速、有效、无创的检查方法,可用于真菌性角膜炎的早期病原学诊断以及治疗观察。     

共焦显微镜在临床诊断真菌性角膜炎中的应用

目的 评价共焦显微镜在临床诊断真菌性角膜炎中的应用价值。方法 用共焦显微镜对52例临床拟诊为真菌性角膜炎的患者进行检查，同时行角膜病灶刮片培养，对两组结果进行比较。结果 综合病史、临床表现及治疗结果，有45例临床诊断为真菌性角膜炎，其中32例角膜刮片培养到真菌，阳性率为71．11％(32／45)；43例共焦显微镜检查观察到真菌菌丝和／或孢子，阳性率为95．56％(43／45)。结论 共焦显微镜可以在活体角膜上直接观察到真菌菌丝，是一种快速、有效和无损伤的检查方法，对真菌性角膜炎的早期诊断、治疗和研究等具有重要的临床价值。     

激光共焦显微镜与角膜组织刮片检查在真菌性角膜炎中应用的比较

目的:对比分析激光共焦显微镜检查与角膜组织刮片检查在真菌性角膜炎诊断中的应用.方法:选择2016-05/08来我院治疗的拟诊为真菌性角膜炎患者77例77眼,同时行激光共焦显微镜检查及角膜组织刮片检查,观察图像特点,对比分析两种方法真菌检出率情况.结果:所有患者中,激光共焦显微镜检出阳性66眼,阴性11眼,检出率为86%;角膜组织刮片检查确诊阳性51眼,阴性26眼,检出率为66%,两者比较差异具有统计学意义(P<0.05).结论:激光角膜共焦显微镜检查的安全性和敏感性相对较高,如果能将激光角膜共焦显微镜检查与角膜组织刮片检查联合应用于临床,对提高真菌性角膜炎患者的诊断率具有重要意义.     

共聚焦显微镜在真菌性角膜炎诊治中的临床应用

目的：评价共聚焦显微镜联合角膜刮片在真菌性角膜炎中的临床应用。

方法：对我院临床诊断为真菌性角膜炎的患者56例56眼行共聚焦显微镜检查和角膜刮片检查,包括涂片后显微镜下检查和真菌培养,并进行药物或联合手术治疗,统计诊断的阳性率和治疗预后。

结果：入院时共聚焦显微镜诊断阳性率为91%,角膜刮片病原学检测诊断阳性率为54%,共聚焦显微镜和角膜刮片联合诊断阳性率为95%。治疗后,单纯药物治疗达到临床治愈者44眼(79%),单纯药物治疗无法控制感染需要进行手术治疗者12眼(21%)。

结论：共聚焦显微镜具有非侵入性、快速、重复性好等特点,联合角膜刮片实验室检查可以提高真菌性角膜炎的诊断阳性率,更好地指导治疗。     

真菌性角膜溃疡刮片前后的共焦显微镜对照研究

目的 对活体真菌性角膜溃疡患者刮片前后的病灶,采用角膜激光共焦显微镜进行观察、比较并总结其特点.方法 取45例(45只眼)角膜溃疡刮片、培养确诊真菌性角膜溃疡的患者,在角膜刮片取材培养前后,采用激光角膜共焦显微镜进行检查,研究其图像特点,并进行对比.结果 在真菌性角膜溃疡刮片取材前,溃疡灶中的真菌菌丝在活体角膜激光共焦显微镜中结构完整、成像清晰.刮片取材后,溃疡灶中的真菌菌丝在活体角膜激光共焦显微镜中结构不完整、成像较模糊,菌丝有断裂.结论 角膜刮片取材会造成真菌性角膜溃疡灶中的真菌菌丝断裂,对真菌性角膜溃疡进行活体共焦显微镜的最佳检查时间应在角膜刮片取材前进行,以提高共焦显微镜对真菌性角膜溃疡的检出水平.     

真菌性角膜炎70例早期临床诊断分析

目的:分析70例真菌性角膜炎早期临床诊断结果,评价共焦激光显微镜在真菌性角膜炎早期临床诊断中的应用价值。方法:用共焦激光角膜显微镜对临床拟诊为真菌性角膜炎的70例患者进行检查,同时行涂片检查真菌菌丝及培养查真菌菌落,对三组结果进行比较。结果:在70例患者中,根据病史、临床表现确诊为真菌性角膜炎,其中共焦激光角膜显微镜的确诊率为94%(66/70),角膜刮片的确诊率为31%(22/70),培养查真菌菌落确诊率为20%(14/70)。激光共焦角膜显微镜检查检出率高于角膜组织刮片(χ2=29.615,P<0.01)及真菌培养(χ2=39.433,P<0.01)检查。结论:共焦激光角膜显微镜是一种无创、快速、有效的活体检查方法,在真菌性角膜炎的早期诊断、治疗和研究中将起重要作用。     

真菌培养和共聚焦显微镜检查诊断非首诊真菌性角膜炎的临床评价

目的：评价真菌培养和共聚焦显微镜检查在非首诊真菌性角膜炎患者诊断中的作用。

方法：采用回顾性研究。对2016-09/2017-02于我院住院治疗的非首诊真菌性角膜炎患者的共聚焦显微镜检查和真菌培养检查进行汇总和分析,利用配对四格表资料的χ²检验统计方法比较两种检查的阳性率。

结果：非首诊真菌性角膜炎患者31例31眼中,真菌培养阳性18眼,阳性率58%,角膜共聚焦显微镜检查阳性6眼,阳性率19%,真菌培养检查的阳性率明显高于共聚焦显微镜检查的阳性率,差异有统计学意义(χ²=7.56,P<0.01)。真菌培养阴性的13例13眼患者中,2眼共聚焦显微镜检查阳性; 共聚焦显微镜检查阴性的25例25眼患者中,14眼真菌培养阳性。

结论：对非首诊真菌性角膜炎患者,真菌培养阳性率较角膜共聚焦显微镜检查阳性率更高。真菌培养是诊断非首诊真菌性角膜炎并明确真菌种属的重要辅助检查方法。在临床诊治过程中,共聚焦显微镜检查因其有着快速、无创、可重复操作的特点,也应作为怀疑真菌性角膜炎患者的首选辅助检查。两种检查方法联合应用可提高诊断阳性率。对于部分辅助检查阴性的患者,不可轻易否定其真菌感染的可能性。     

角膜溃疡清创术联合抗真菌药物治疗真菌性角膜炎

目的:总结角膜溃疡清创术联合抗真菌药物治疗真菌性角膜炎的效果。方法:收集2010-09/2011-09在我院就诊的真菌性角膜炎患者30例,对患者进行常规角膜刮片检查,并对全部患者施行角膜溃疡清创术,根据角膜炎的严重程度联合给予氟康唑或那他霉素眼水点眼治疗,清创术治疗无效者采用自体结膜覆盖术及板层角膜移植术治疗,随访观察治疗效果。结果:角膜刮片阳性结果 27例,阳性率90%,2例于外院行共聚焦显微镜检查菌丝阳性;1例为明确植物外伤史的8岁儿童,不能配合真菌检查。轻度角膜炎6例,施行角膜溃疡清创术联合氟康唑眼水点眼治疗全部痊愈,病程2～3wk;中度度角膜炎17例,行角膜溃疡清创术联合那他霉素眼水点眼治疗痊愈12例,病程4～8wk;重度角膜炎7例,行角膜溃疡清创术联合那他霉素眼水点眼治疗痊愈3例,病程8～10wk。角膜溃疡清创术联合抗真菌药物治疗真菌性角膜炎有效率70%。结论:角膜溃疡清创术联合抗真菌药物治疗真菌性角膜炎操作简单,是一种经济、有效、可行的治疗方式。     

共焦激光角膜显微镜在真菌性角膜炎诊断中的应用

目的评价共焦激光角膜显微镜在临床诊断真菌性角膜炎中的应用价值。方法用共焦激光角膜显微镜对临床拟诊为真菌性角膜炎的26例患者进行检查,同时行涂片检查真菌菌丝,对两组结果进行比较。结果26例患者中,根据病史、临床表现24例确诊为真菌性角膜炎,其中共焦激光角膜显微镜的确诊率为95.83%(23/24),角膜刮片的确诊率为83.33%(20/24)。结论共焦激光角膜显微镜是一种快速、有效、无损伤的活体检查方法,在真菌性角膜炎的早期诊断、治疗和研究中将起重要作用。     

应用激光共焦显微镜诊断真菌性角膜炎的临床研究

目的:应用激光共焦显微镜观察真菌性角膜炎患者图像特点及菌丝和孢子检出率,探讨激光共焦显微镜检查在真菌性角膜炎临床诊断中的意义。方法:对41例41眼经门诊确诊为真菌性角膜炎的患者行角膜激光共焦显微镜检查,观察不同治疗期真菌性角膜炎患者活体角膜各层图像特点。结果:共焦显微镜下真菌性角膜炎患者图像有如下特点:(1)病变部位角膜各层形态结构破坏明显;角膜上皮至基质不同程度水肿;炎细胞浸润;神经结构破坏;基质层结构紊乱,透过度降低;(2)真菌菌丝是本病的特异性诊断依据,不同菌种感染在镜下菌丝有不同的影像学特点;(3)不同病变时期和治疗阶段,图像有很大差异,真菌菌丝的检出并不是诊断本病的唯一依据。结论:激光共焦显微镜检查具有无创、及时等优点,在临床诊断真菌性角膜炎中有重要参考意义,尤其是指导临床早期诊断、合理治疗及评价预后。     

Sensitivity and specificity of confocal scan in the diagnosis of infectious keratitis

PURPOSE: To compare the sensitivity and specificity of confocal scan with the results of corneal and/or contact lens case smear and culture in the diagnosis of infectious keratitis. METHODS: This was a diagnostic test study. Confocal scan and corneal and/or contact lens case smear and culture were performed in all patients with a clinical diagnosis of infectious keratitis at Labbafinejad Medical Center from 2004 to 2006. RESULTS: A total of 133 eyes of 133 patients with a mean age of 48.0 +/- 22.6 years (range, 9-83 years) were included in the study. Sixty-nine (51.9%) cases were men, and 64 (48.1%) were women. Twenty-eight cases (21%) were contact lens wearers; history of trauma and previous ocular surgery was present in 21% and 38.3%, respectively. Corneal and/or contact lens case smear and culture were positive in 71 eyes (53.4%), including 40 cases of bacterial, 16 cases of fungal, and 15 cases of acanthamoeba infection. Confocal scan was positive in 50 cases (37.6%), which revealed hyphae-like structures in 27 cases and cyst and/or trophozoite-like structures in 23 cases. The sensitivity and specificity of confocal scans were 100% and 84% for the diagnosis of acanthamoeba keratitis versus 94% and 78% for fungal keratitis, respectively. CONCLUSIONS: In vivo corneal confocal scan is a rapid noninvasive tool for the diagnosis of acanthamoeba and fungal keratitis with high sensitivity and specificity compared with smear and culture. It may also be helpful in excluding fungal or acanthamoeba-like structures in cases with negative bacteriologic results and in early bacterial keratitis before clarification of microbiologic results.     

实验室诊断技术在真菌性角膜炎诊断敏感性的对比研究

目的：分析真菌涂片、真菌培养和病理检查在诊断真菌性角膜炎上的相关性及临床意义。
　　方法：收集2012-01/2014-12诊断为真菌性角膜炎的患者110例110眼,对真菌涂片、真菌培养及病理检查结果进行回顾性分析总结。真菌涂片行角膜刮片,用10%氢氧化钾制成湿片镜检,同时行革兰染色镜检;真菌培养采用马铃薯葡萄糖琼脂培养基( PDA)培养;病理检查的标本为角膜移植切除手术标本,采用石蜡切片,使用苏木素-伊红( HE)、六胺银、过碘酸-雪夫( PAS)染色镜检。
　　结果：真菌涂片阳性者50例(45.5%);真菌培养阳性者55例(50.0%);病理检查阳性者88例(80.0%)。真菌涂片与病理检查均为阳性者50例,真菌涂片与病理检查均阴性者22例,真菌涂片与病理检查的符合率为65.5%。真菌培养与病理检查均为阳性者55例,真菌培养与病理检查均阴性者22例,真菌培养与病理检查的符合率为70 .0%。真菌涂片结果阴性者60例中,有38例经病理检查确诊为阳性,占63.3%;真菌培养结果阴性者55例中,有33例经病理检查确诊为阳性,占60 .0%。
　　结论：病理检查的敏感性最好,真菌涂片、真菌培养及病理检查联合应用能够提高真菌性角膜炎的诊断水平,降低漏诊、误诊。     

Clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis

AIM: To study the clinical observation of removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis. METHODS：A retrospective study was done to 10 patients (10 eyes) who had accepted removal of the necrotic corneal tissue combined with conjunctival flap covering surgery for fungal keratitis,the diagnosis by corneal scraping and smear examination or confocal microscopy check hyphae.Local and systemic antifungal therapy more than one week for all patients, corneal ulcer enlarge or no shrink.Slit lamp microscope examination the diameter of corneal ulcer about 2mm-4mm.Anterior segment optical coherence tomography（AS-OCT）examine the depth of corneal ulcer between 1/3-1/2,infiltrate corneal stroma about 20um-80um,the diameter of corneal ulcer about 3mm-6mm.Type-B ultrasonic exclusion endophthalmitis. Complete removal lesions until transparent of stroma, make conjunctival flap equal or greater than ulcer 1mm nearby conjunctiva. Continued antifungal therapy. The vision, fungal recurrence, conjunctival flap rollback or desquamate were analysed. RESULTS：Ten patients had success done this surgery, the corneal ulcer was not enlarge and healing afteroperation.7 cases were bridging conjunctival flap and 3cases were single conjunctival flap. Preoperation vision above 0.1 had 8 cases,7 cases had vision above 0.1 one week after surgery, while 1 cases vision droped from 0.3 to 0.05.There was not recurrent for fungal,2 cases conjunctival flap rollback:1 case was bridging and 1case was single flap, no conjunctival flap desquamate. CONCLUSION：It is safe and effective to perform removal of the necrotic corneal tissue combined with conjunctival flap covering surgery under the guidance of the AS-OCT in treatment of fungal keratitis which werenot sensitive or aggravate for antifungal drugs.     

Antifungal chemotherapy for fungal keratitis guided by in vivo confocal microscopy

Background To evaluate antifungal chemotherapy in patients with fungal keratitis guided by in vivo confocal microscopy. Methods A total of 121 patients (121 eyes) with fungal keratitis were enrolled in this study. Confocal microscopy was performed in real time after topical and/or oral antifungal chemotherapy. Hyphal density and morphology, composition of inflammatory cells, and appearance of corneal stromal cells at the central and peripheral corneal lesions were recorded. Antifungal therapy discontinued at 1 week after hyphae and inflammatory cells could not be detected, and affected corneal stromal cells became visible. Results Successful outcomes were achieved in 110 patients (90.9%). By confocal microscopy, we observed the gradual decrease of hyphae-positive sites and hyphal density during the chemotherapy. The inflammatory cells reduced in number and heterogeneity, while corneal stromal cells recovered. The antifungal drugs were tapered according to the changes in hyphae, inflammatory cells, and corneal stromal cells. There was no fungal recurrence during the 2-month follow-up period. The other 11 patients (9.1%) had deteriorated infection within 1 week of antifungal therapy, and therefore were subjected to corneal transplantation. Conclusions In vivo confocal microscopy appears to be an effective approach to guide antifungal chemotherapy. It allows comprehensive evaluation of hyphae, inflammatory cells, and corneal stromal cells in real time, and provides valuable and objective information required in selecting and adjusting therapeutic regimens for the treatment of fungal keratitis.     

Evaluation of impression smear in the diagnosis of fungal keratitis

PURPOSE: To evaluate and compare impression smear with the conventional mechanical corneal scraping by potassium hydroxide (KOH) method in the diagnosis of fungal keratitis. MATERIALS AND METHODS: Fifty consecutive cases of clinically suspected fungal keratitis were included. Under topical anesthesia, impression smear was obtained from the lesion, followed by the conventional mechanical scraping and both were transferred to sterile glass slides. The slides were examined under direct microscopy with KOH 10% wet mount. Impression smear KOH was compared with the scraping KOH and culture. RESULTS: The KOH smear was positive in 36 (72%) patients with impression smear technique and positive in 35 (70%) patients from corneal scrapings. Sensitivity of impression smear technique as compared to scraping was 97.14%, specificity 92.86%, positive predictive value 94.4%, negative predictive value 92.86%, false positive rate 5.56%, false negative rate 7.14% and accuracy was 94%. As compared to culture, the sensitivity was 100%, specificity 46.67%, positive predictive value 55.56%, negative predictive value 100%, false positive rate 53.33%, false negative rate 0% and accuracy was 68%. CONCLUSION: Impression smear KOH examination is comparable to conventional mechanical corneal scraping KOH examination in making a tentative diagnosis of fungal keratitis and can be accurately relied upon for initiating anti-fungal therapy.     

Detection of <Emphasis Type="Italic">Aspergillus</Emphasis> keratitis in ocular infections by culture and molecular method

The incidence of fungal keratitis has increased in recent years. Since accurate and rapid diagnosis is important for the proper management of this infection, we evaluated the use of nested PCR to detect Aspergillus-specific DNA in corneal scrapings for the diagnosis of Aspergillus keratitis. From April 2005 to September 2008, patients with eye findings suspicious for fungal keratitis were enrolled for corneal sampling. After ocular examination, we carried out laboratory testing including direct microscopy and culture examination of corneal scrapings, nested PCR and histopathological examination of the corneal tissues obtained at surgery. Of the 57 infected eyes suspicious for fungal keratitis, 32 (56.14%) were reported as Aspergillus keratitis by PCR assay, from which 12 (37.5%) were positive in culture and direct smear for Aspergillus keratitis, and 20 (62.5%) had clinical signs of fungal keratitis (probable). All PCR-positive cases were judged to have fungal infections, based on their responses to the antifungal treatment. The use of molecular methods in high-risk patients can be helpful in diagnosis of fungal infections and can have important public health implications for both the treatment and prevention of suppurative corneal ulcerations. The significance of this study lies in the analysis of the proven cases in terms of both laboratory findings and responses to the treatment.