Novati R, Castagna A, Morsica G, Vago L, Tambussi G, Ghezzi S, Gervasoni C, Bisson C, d'Arminio Monforte A, Lazzarin A
Infectious Diseases Department, San Raffaele Scientific Institute, Milan, Italy.
AIDS. 1994 Dec;8(12):1691-4. doi: 10.1097/00002030-199412000-00008.
To study the accuracy of polymerase chain reaction (PCR) for Toxoplasma gondii DNA in the cerebrospinal fluid (CSF) of AIDS patients for the diagnosis of T. gondii encephalitis.
Eighty-two AIDS patients with brain lesions. At autopsy, 19 patients (group A) had toxoplasmic encephalitis and 33 (group B) primary brain lymphoma or other infections. Brain histology was not available for 30 patients; cerebral lesions improved after anti-Toxoplasma therapy in 16 (group C), but there was no improvement in 14 patients (group D).
T. gondii RH strain was serially diluted in microplate wells. After heat denaturation, nested PCR was performed on diluted tachyzoites and on 10 microliters CSF with primers flanking the B1 repetitive region of T. gondii genome.
DNA from one to five tachyzoites was detected in each experiment. PCR was positive in eight (42.1%) out of 19 group A samples, none of the group B samples, 10 (62.5%) out of 16 group C samples and none of the group D samples. Among group A and C patients, PCR was positive in all 11, and in seven out of 24 (29.1%; P < 0.04) patients who had received anti-Toxoplasma therapy for less or more than 1 week at the time of rachicentesis, respectively.
Nested PCR for T. gondii in CSF may improve early differential diagnosis of AIDS-associated focal brain lesions. Higher diagnostic accuracy was achieved when lumbar puncture was performed in the first week of anti-Toxoplasma therapy.
研究聚合酶链反应（PCR）检测艾滋病患者脑脊液（CSF）中弓形虫DNA以诊断弓形虫脑炎的准确性。
82例有脑部病变的艾滋病患者。尸检时，19例患者（A组）患有弓形虫脑炎，33例（B组）患有原发性脑淋巴瘤或其他感染。30例患者无法获得脑活检组织；16例患者（C组）经抗弓形虫治疗后脑部病变改善，14例患者（D组）无改善。
将弓形虫RH株在微孔板孔中进行系列稀释。热变性后，使用位于弓形虫基因组B1重复区域两侧的引物，对稀释后的速殖子和10微升脑脊液进行巢式PCR。
每次实验均检测到1至5个速殖子的DNA。A组19个样本中有8个（42.1%）PCR呈阳性，B组样本均为阴性，C组16个样本中有10个（62.5%）呈阳性，D组样本均为阴性。在A组和C组患者中，在腰穿时分别接受抗弓形虫治疗少于或多于1周的所有11例患者以及24例患者中的7例（29.1%；P<0.04）中PCR呈阳性。
脑脊液中弓形虫的巢式PCR可能改善艾滋病相关局灶性脑病变的早期鉴别诊断。在抗弓形虫治疗的第一周进行腰穿时诊断准确性更高。
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