Rapid detection of cytomegalovirus DNA in cerebrospinal fluid of AIDS patients with neurologic disorders.

A polymerase chain reaction (PCR)-based method was used to detect cytomegalovirus (CMV) DNA in 82 cerebrospinal fluid (CSF) samples from 67 patients infected by human immunodeficiency virus (HIV). The test was positive for 14 patients, 8 of whom had CMV-related neurologic disease proven by viral culture of CSF or histologic examination. Encephalitis was the most frequent manifestation in patients with positive PCR results, but CMV DNA was also present in some patients with peripheral neuropathy or polyradiculomyelitis. All patients with proven CMV neurologic disease were positive by PCR. In contrast, viral culture was negative for 4 of the 8 patients and pathologic studies were available only for 5. The specificity of the PCR-based assay could not be assessed precisely because of the lack of a reference standard, but the results correlated well with clinical course and results of the other methods. These findings suggest that the PCR-based method may be a useful noninvasive tool for the rapid diagnosis of CMV-related neurologic disease.     

Rapid diagnosis of cytomegalovirus infection in congenital neonates

Immunostaining was compared with PCR for diagnosis of congenital CMV infection. IgM and IgG antibody assays were also performed in parallel. Immunostaining gave sensitivity and specificity of 60% and 97% respectively. Correlations among immunostaining, PCR and the presence of IgM antibody was reported. Immunostaining can be used for early diagnosis of congenital CMV infection in parallel with detection of IgM antibody.     

AIDS合并巨细胞病毒脑炎临床分析

目的探讨艾滋病（AIDS）合并巨细胞病毒脑炎（CMVE）的临床特点及治疗转归。方法回顾性地收集北京地坛医院2011年3月-2013年2月收治的8名合并CMVE的AIDS病人的临床资料,分析其症状和体征、辅助检查、病毒学检查及治疗转归情况。结果 8例病人脑脊液CMV脱氧核糖核酸（DNA）或CMV-IgM抗体阳性,但血清中CMV DNA可阴性。7例CD4＋淋巴细胞计数均〈30/μL,轻症、早期病人可不伴随神经精神症状,头颅核磁共振（MRI）检查可无异常,7例患者经抗CMV及高效抗反转录病毒治疗（HAART）后均获得临床好转。结论合并CMVE易出现在CD4＋淋巴细胞计数低的AIDS病人中,及时对可疑病人进行脑脊液CMV DNA或CMV-IgM抗体检查,有利于早期诊断和治疗。     

Prevalence of human cytomegalovirus (HCMV) gB genotypes in Thai patients

Human cytomegalovirus (HCMV) infection can cause asymptomatic to symptomic diseases leading to morbidity and mortality especially in immunocompromized patients. One factor of the difference in clinical outcome is the distinction of HCMV strain. As HCMV glycoprotein (g)B plays an important role in viral entry and neutralizing antibody induction, HCMV gB genotypes were determined in 161 clinical specimens containing HCMV-DNA obtained from patients at King Chulalongkorn Memorial Hospital, Bangkok, Thailand during the year 2000 and 2004. Of the 113 (70%) samples that were able to be genotyped, mixed gB genotype was demonstrated in 35%, followed by gB1 (33%), gB3 (15%), gB2 (11%), and untyped (7%); gB4 was not detected. The distribution of HCMV gB genotypes between genders was not significantly different. Mixed gB genotype (35%) was found in HIV- infected patients.     

Anal ulcerations due to cytomegalovirus in patients with AIDS

Lesions due to cytomegalovirus (CMV) are frequent in the immunocompromised patient. This is particularly the case in patients with AIDS, where the colon and rectum are the regions most often involved. The authors report six cases of anal ulcerations due to CMV in patients with AIDS. These lesions, confirmed histologically, were either isolated or associated with other localizations. Treatment is based on specific antiviral agents but resistance and recurrence can occur. The current report underlines the importance of histologic examination of anal lesions in HIV-infected subjects.     

20例AIDS合并巨细胞病毒脑炎的临床分析

目的探讨AIDS并发巨细胞病毒脑炎的临床特点、诊治及转归。方法回顾性分析北京地坛医院2012年1月—2015年1月明确诊断为AIDS并发巨细胞病毒脑炎的20例患者临床资料,分析其临床表现特点、脑脊液检测结果及诊治过程。结果 20例患者均处于AIDS晚期的严重免疫缺陷状态,临床表现不典型,多有智力下降、头痛、意识障碍以及神经功能受损表现,影像学检查无显著特异性,疗效较差。结论巨细胞病毒脑炎临床表现缺乏特异性,须尽早确诊治疗以降低病死率。     

45例AIDS合并巨细胞病毒性视网膜炎临床特点分析

目的探究AIDS合并巨细胞病毒(cytomegalovirus,CMV)性视网膜炎临床诊断及治疗措施。方法收集2012年2月—2014年2月收治的45例AIDS合并CMV性视网膜炎患者临床资料并进行回顾性分析,分析患者一般资料、临床特征、实验室检查结果、治疗及转归。结果患者眼部常见的症状为视力下降和视物模糊,分别为41例(91.11%)和23例(51.11%),另有3例(6.67%)发生失明。眼底检查发现视网膜血管炎和视神经炎,分别为31例(68.89%)和5例(11.11%),另有5例(11.11%)发生视网膜脱离。患者CD4+T细胞计数平均为(56.3±12.6)个/μl,CMV-Ig M阳性15例(33.33%),CMV抗原阳性18例(40.00%)。应用替诺福韦+拉米夫定+依非韦伦进行抗病毒治疗后,19例治愈(42.22%),19例好转(42.22%),7例无效(15.56%)。结论 AIDS患者免疫功能缺损,易合并眼部CMV感染造成视网膜炎症,临床以视力下降和视力模糊为主,眼底检查常见为视网膜血管炎和视神经炎,积极治疗可获得显著效果。     

艾滋病相关性弓形虫脑炎的影像学诊断

目的 评估艾滋病脑弓形虫脑炎的影像表现的诊断价值. 方法 回顾分析17例艾滋病脑弓形虫脑炎患者的影像表现. 结果 病变多为双侧多发,可少发;侵犯基底核12例,丘脑3例,皮髓质连接区1例;累及小脑及脑干1例;CT呈低密度,MRI呈长T1、长T2信号;周围水肿效应明显;增强扫描,小环状、螺旋状或靶形增强12例;巨大环形增强1例;多发病灶15例,单发2例. 结论 位于基底节区的多发小环状、螺旋状或靶形增强,高度提示脑弓形虫脑炎.CT和MRI对弓形虫脑炎是有效的诊断方法.MRI对病灶检出率明显高于CT.     

Intestinal perforation due to cytomegalovirus infection in patients with AIDS

Intestinal perforation due to cytomegalovirus (CMV) infection in patients with AIDS is the most common life-threatening condition requiring emergency celiotomy in these patients. The authors describe a patient with AIDS with intestinal perforation due to CMV infection, and review 14 additional cases reported in the English-language surgical literature. The diagnostic triad of pneumoperitoneum on x-ray, evidence or history of CMV infection, and AIDS occurred in 70 percent of patients. The most common site of intestinal perforation was the colon (53 percent), followed in frequency by the distal ileum (40 percent) and appendix (7 percent); perforation usually occurred between the distal ileum and splenic flexure of the colon. Colonoscopy, rather than sigmoidoscopy, is recommended as a screening examination in patients with AIDS suspected of having colonic uleration due to CMV infection. Multiple biopsies of ulcerated tissue should be obtained. Gross and microscopic analyses of involved intestinal tissue reveal the characteristic findings of ulceration and CMV infection. Despite aggressive therapy, the operative mortality rate in patients with AIDS with intestinal perforation due to CMV infection was 54 percent and the overall mortality rate was 87 percent. Postoperative complications occurred in most patients and consisted mainly of systemic sepsis and pneumonia caused by Pneumocystis carinii infection. An increased awareness of this syndrome by physicians frequently called on to manage patients with AIDS is recommended.     

Sonographic diagnosis of cholangitis in AIDS patients

Eleven acquired immunodeficiency syndrome (AIDS) patients were initially evaluated by ultrasound of the liver because of biochemical evidence of cholestasis. Sonography was very useful for demonstrating dilatation and/or wall thickening of the common bile duct associated with dilated intrahepatic bile ducts. Periportal hypo- or hyperchoic areas were also visualized. In seven patients, gallbladder walls were thickened. In six patients, the diagnosis of cholangitis was confirmed by endoscopic retrograde cholangiography and/or histology. Cytomegalovirus infection or digestive tract cryptosporidiosis were often present and accounted for diarrhea in most patients. We believe the association of upper right quadrant pain and anicteric cholestasis in the course of AIDS is suggestive of cholangitis, but a disparity may exist between mild symptoms and sonographically demonstrated extensive biliary involvement.     

Gastrointestinal cytomegalovirus manifestations in AIDS

Cytomegalovirus (CMV) infection is one of the most important intestinal opportunistic infections in AIDS. In severe cases ulcerations and colitis are the commonest manifestations. 184 HIV positive patients with gastrointestinal symptoms were investigated by endoscopy of the gastrointestinal tract. While culture, immunohistochemical staining and histology from biopsies were performed, the results of all three methods were compared. In one third the cases CMV associated lesions could be found by endoscopy. Erosions or ulcerations are the most frequent tissue lesions. In 95% the culture was positive. In addition, immunohistochemical staining in 75% and histology in 61.7% were positive in patients with more serious manifestations. For early diagnosis endoscopy of the gastrointestinal tract and histological, histochemical and microbiological investigations of biopsies are essential.     

Foscarnet therapy for ganciclovir-resistant cytomegalovirus retinitis in patients with AIDS.

Infections caused by cytomegalovirus (CMV) resistant in vitro to ganciclovir, defined as requiring greater than 6 mumols of ganciclovir for ED50 have developed in some AIDS patients with progressive CMV retinitis despite chronic ganciclovir therapy. Two such patients (CMV isolates ED50, 9.5-14.5 mumols) were treated with foscarnet, an antiviral pyrophosphate analogue to which both patients' isolates demonstrated in vitro susceptibility (ED50, less than or equal to 300 mumols). Each patient had documented retinitis progression, at 2- and 1- to 5-week intervals, respectively, despite high-dose intravenous ganciclovir therapy. Both patients responded to foscarnet therapy with cessation of viral shedding in urine and blood. After foscarnet therapy was started, retinitis stabilized in the two patients for 12 and 25 weeks, respectively, before progression recurred. Therefore, foscarnet may be effective in immunocompromised patients with rapidly progressive CMV retinitis whose CMV isolates have developed in vitro resistance to ganciclovir.     

Effect of highly active antiretroviral therapy on cytomegalovirus antigenaemia in AIDS patients

To assess the effect of highly active antiretroviral therapy (HAART) on cytomegalovirus (CMV) antigenaemia in AIDS patients, 70 patients with CD4+ cell counts < or = 50/mm3 and positive anti-(CMV) immunoglobulin G (IgG) were tested at 15-30 day intervals for CMV antigenaemia. We selected those patients who had been followed up for more than 3 months. Three patient profiles were defined: A, followed up before the introduction of HAART; B, followed up before and after the use of HAART; and C, followed up after the use of HAART. Thirty-nine patients were included, 12 in group A, 17 in group B, and 10 in group C. Group A patients presented a lower median CD4+ cell count compared with groups B and C patients (9, 122 and 127 cells/mm3, respectively), with the increase in the last 2 groups being related to the use of HAART (P<0.001). A lower proportion of positive antigenaemia was observed in group B after the introduction of HAART compared with the time before HAART (P=0.02). HAART caused an immunological improvement and was found to be associated with negativity of CMV antigenaemia.     

The diagnosis of anal ulcers in AIDS patients

Recent reports have suggested that routine microscopic evaluation of anal ulcer tissue from AIDS patients is not the most accurate way to diagnose viral infection. This study was undertaken to determine if either viral culture (VC) or immunohistochemistry (IHC) can improve the diagnostic accuracy as compared with routine hematoxylin and eosin (H&E) staining. Specifically, we sought to identify inclusion bodies of cytomegalovirus (CMV) or herpes simplex virus (HSV) to assist in the diagnosis of CMV or HSV. All patients had clinical evidence of an anal ulcer or a nonhealing anal fissure. Duration of symptoms ranged from 1 week to 3 months with a mean of 6 weeks. All specimens were submitted for viral culture in addition to routine H&E staining; immunohistochemistry was also performed. Twenty-five paraffin-embedded anal ulcer biopsies from 23 male patients (age range 27–73; mean 37.4 years) with the diagnosis of AIDS or AIDS-related complex (ARC) were reviewed over a 4 year period (1988–1992). Routine H&E staining revealed 6 (22%) specimens with CMV inclusions. Four of these 6 reacted positively with IHC (67%) and one was positive on viral culture (17%). In the remaining 19 specimens that did not reveal infection with CMV (78%), IHC was positive in 2 patients (10%) and viral culture was positive in 1 patient (5%). Although HSV was not seen in any of the specimens on H&E staining, IHC was positive in one patient (3.5%) and viral culture reacted positively in 8 (29%) specimens. Thus IHC is a good confirmatory test for CMV inclusions and can be used to achieve a definitive diagnosis in equivocal cases. However, neither viral culture nor IHC can replace the traditional careful pathologist in the detection of viral inclusions.

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Résumé Des publications récentes ont suggéré que l'examen microscopique de tissu prélevé dans des ulcères anaux chez des patients atteints de SIDA n'était pas une méthode sûre de diagnostic de l'infection virale. Cette étude a été entreprise pour déterminer si les cultures virales (VC) ou les examens immuno-histochimiques (IHC) peuvent améliorer la sécurité du diagnostic en comparaison avec les colorations usuelles à l'hématoxyline et éosine (H&E). Spécifiquement, nous souhaitions identifier des inclusions virales de cytomégalovirus (CMV) ou d'herpès symplex (HSV) pour assurer un diagnostic d'infection à CMV ou HSV. Tous les patients étaient porteurs d'ulcères anaux cliniquement manifestes ou de fissures anales non cicatrisées. La durée des symtômes variait d'une semaine à trois mois avec une moyenne de six semaines. Tous les prélèvements ont fait l'objet de culture virale en complément à la coloration habituelle à l'hématoxyline éosine; des examens immuno-histo-chimiques ont également été réalisés. Vingt-cinq biopsies d'ulcère anal enrobées dans de la paraffine prélevées chez 23 patients mâles (ägés de 27 à 73 ans avec une moyenne de 37,4 ans) atteints de SIDA ou de ARC ont été revus sur une période de 4 ans (1988–1992). L'examen histologique avec coloration à l'hématoxyline éosine a montré chez 6 malades (22%) des inclusions à CMV. Quatre de ces 6 patients ont réagi positivement à un dosage immuno-histochimique (67%) et un s'est révélé positif à la culture virale (17%). Dans les 19 specimens restants qui n'ont pas révélé d'infection à CMV (78%), l'immuno-histo-chimie a été positive chez deux patients (10%) et la culture virale était positive chez un patient (5%). Bien qu'une infection à HSV n'ait pas été mise en évidence dans les prélèvements colorés à l'hemotoxyline éosine, l'immuno-histo-chimie s'est révélée positive chez un malade (3,5%) et la culture virale s'est révélée positive chez 8 patients (29%). L'immuno-histo-chimie constitue donc un bon test de confirmation d'inclusion à CMV et peut être utilisée pour assurer un diagnostic définitif dans des cas douteux. Cependant, ni la culture virale ni l'immuno-histo-chimie ne peuvent remplacer le pathologue soigneux au cours de la recherche d'inclusion virale.

     

Laboratory diagnosis of cytomegalovirus infection and disease in immunocompromised patients

PURPOSE OF REVIEW: To review new developments in PCR technology as they apply to detecting cytomegalovirus viremia and pneumonia, recent advances in detecting CMV resistance to antivirals and assays of specific CMV lymphocyte function. RECENT FINDINGS: This review summarizes the attempts to use real time PCR for cytomegalovirus deoxyribonucleic acidemia and to compare it to conventional PCR and antigenemia, it also reviews the use of quantitative PCR on bronchoalveolar lavage to assist in the diagnosis of CMV pneumonia.Phenotypic assays of susceptibility in tissue culture are much too slow to assist clinical decisions, taking weeks for completion. Genotypic assays may be performed directly on clinical samples such as blood, and cerebrospinal fluid and can be done by sequencing in a very few days.Finally, assays of lymphocytic functional responsiveness to cytomegalovirus can be used to identify transplant recipients at continuing risk for cytomegalovirus disease. SUMMARY: Assays for CMV DNA or antigen in blood are superior to culture for documenting viremia and pneumonia. Genotypic assays have largely replaced phenotypic assays for CMV resistance to antivirals. Lymphocyte responses to CMV antigen(s) may identify patients at risk for CMV disease.     

Cytomegalovirus encephalitis in two patients with AIDS receiving ganciclovir for cytomegalovirus retinitis

Cytomegalovirus (CMV) encephalitis has been reported with increasing frequency in patients with AIDS. Nevertheless, the management of CMV-related encephalitis appears to be problematic and data in the literature on the clinical efficacy of ganciclovir therapy is sparse and controversial. We describe two patients with AIDS who developed CMV encephalitis while receiving ganciclovir maintenance therapy for CMV retinitis. Moreover, there was no improvement in neurological status or virological and radiological response during a further induction course of ganciclovir. These observations suggest that the currently recommended therapeutic protocols with ganciclovir are not effective in the prevention and treatment of CMV encephalitis in patients with AIDS.