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两例嗜血性分枝杆菌感染所致皮肤溃疡的诊断:通过聚合酶链反应-限制性内切酶分析在临床样本中进行直接鉴定

The diagnosis of two cases of cutaneous ulcer caused by infection with Mycobacterium haemophilum: direct identification in a clinical sample by polymerase chain reaction-restriction endonuclease analysis.

作者信息

Da Mata Omaira, Pérez Alfonzo Ricardo, Natera Ivelisse, Sucre Rosibel del Carmen, Bello Teresita, de Waard Jacobus H

机构信息

Laboratorio de Tuberculosis and Centro Clínico de Dermatología y Enfermedades Tropicales, Instituto de Biomedicina, Universidad Central de Venezuela, Caracas.

出版信息

Int J Dermatol. 2008 Aug;47(8):820-3. doi: 10.1111/j.1365-4632.2008.03702.x.

Abstract

BACKGROUND

Mycobacterium haemophilum was first recovered from subcutaneous lesions of a patient with Hodgkin's disease. Because of its special growth requirements (it grows at 30-32 degrees C and requires iron-supplemented medium), the organism cannot be isolated using routine culture techniques for other mycobacteria. Only a few developed countries have reported infection with this mycobacterium. We report the first two cases diagnosed in Venezuela.

METHODS

The diagnosis of the first case was established using polymerase chain reaction (PCR)-restriction endonuclease analysis of the gene encoding the 65-kDa heat shock protein (hsp65) for the direct identification of M. haemophilum in a clinical specimen in which bacilli were observed on acid-fast smear, but growth was not detected by standard culture procedures.

RESULTS

After recognizing this bacterium as a possible cause of infection in our setting, clinical samples of cutaneous lesions were routinely cultured on blood agar at 30 degrees C for at least 6 weeks, which resulted in the diagnosis of the second case.

CONCLUSIONS

Dermatologists should consider this bacterium in immunocompromised patients with cutaneous ulcerating lesions. Material from the lesions can be screened for mycobacteria using an acid-fast stain and, if acid-fast bacilli are seen, PCR analysis of mycobacterial hsp65 can be an effective tool for early diagnosis. Appropriate culture methods are required for bacteriologic confirmation of infection with M. haemophilum.

摘要

背景

嗜血性分枝杆菌最初是从一名霍奇金病患者的皮下病变中分离出来的。由于其特殊的生长要求(在30 - 32摄氏度生长且需要补充铁的培养基),无法使用针对其他分枝杆菌的常规培养技术分离该菌。只有少数发达国家报道过这种分枝杆菌感染。我们报告委内瑞拉诊断出的首例两例病例。

方法

首例病例的诊断是通过对编码65 kDa热休克蛋白(hsp65)的基因进行聚合酶链反应(PCR)-限制性内切酶分析来直接鉴定临床标本中的嗜血性分枝杆菌,该临床标本在抗酸涂片上观察到杆菌,但标准培养程序未检测到生长。

结果

在认识到这种细菌可能是我们地区感染的病因后,对皮肤病变的临床标本常规在30摄氏度的血琼脂上培养至少6周,从而诊断出第二例病例。

结论

皮肤科医生应在患有皮肤溃疡性病变的免疫功能低下患者中考虑这种细菌。可使用抗酸染色对病变材料进行分枝杆菌筛查,如果看到抗酸杆菌,分枝杆菌hsp65的PCR分析可以是早期诊断的有效工具。需要适当的培养方法来对嗜血性分枝杆菌感染进行细菌学确认。

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