Juan Agapito

Clinical Evaluation of a 16S Ribosomal RNA Polymerase Chain Reaction Test for the Diagnosis of Lymph Node Tuberculosis

UNLABELLED Reports on the sensitivity of polymerase chain reaction (PCR) for the diagnosis of lymph node tuberculosis (TB) show divergent results. We evaluated the accuracy of the Roche Amplicor Mycobacterium tuberculosis PCR test with lymph node aspirate and biopsy samples. METHODS The study was conducted at a public reference hospital in Lima, Peru. From the period of January 2003 to January 2004, we included patients who had lymphadenopathy and in whom the attending physician suspected TB. Aspirate and biopsy samples were submitted for culturing in Lowenstein-Jensen medium, for histopathologic testing, and for PCR. The sensitivity and specificity of PCR were calculated against a reference standard based on histopathologic findings and culture. RESULTS Our study included 154 patients. Median age was 29 years (interquartile range, 21-40 years); 97 patients (62.9%) were men. Twenty-nine patients (18.8%) had acid fast bacilli-positive histopathologic findings, and 44 (28.6%) had a positive culture result. Using the combination of histopathologic findings and culture as reference standard, 55 patients (35.7%) had a diagnosis of tuberculous lymphadenitis. The sensitivity of the PCR test was 58.2%, and the specificity was 93.9%. For biopsy tissue only, the sensitivity of PCR was 52.7%, and the specificity was 97.0%. For aspirate samples only, the sensitivity of PCR was 47.3%, and the specificity was 96.0%. CONCLUSION The Amplicor PCR test revealed low sensitivity and high specificity for the diagnosis of lymph node TB. The sensitivity was higher in cases in which the bacillary load was high--in acid fast bacilli-positive samples and among HIV-infected patients. Considering the results of microbiological and PCR tests together, there was still a patient group in whom no final diagnosis could be established.

Novel hypertonic saline–sodium hydroxide (HS–SH) method for decontamination and concentration of sputum samples for Mycobacterium tuberculosis microscopy and culture

This study evaluated a new decontamination and concentration (DC) method for sputum microscopy and culture. Sputum samples from patients with suspected pulmonary tuberculosis (TB) (n=106) were tested using the proposed hypertonic saline-sodium hydroxide (HS-SH) DC method, the recommended N-acetyl-L-cysteine-sodium citrate-sodium hydroxide (NALC-NaOH) DC method and unconcentrated direct smear (Ziehl-Neelsen) techniques for the presence of mycobacteria using Löwenstein-Jensen culture and light microscopy. Of 94 valid specimens, 21 (22.3%) were positive in culture and were further characterized as Mycobacterium tuberculosis. The sensitivity for acid-fast bacilli (AFB) smears was increased from 28.6% using the direct method to 71.4% (HS-SH) and 66.7% (NALC-NaOH) using DC methods. Both concentration techniques were highly comparable for culture (kappa=0.794) and smear (kappa=0.631) for AFB. Thus the proposed HS-SH DC method improved the sensitivity of AFB microscopy compared with a routine unconcentrated direct smear; its performance was comparable to that of the NALC-NaOH DC method for AFB smears and culture, but it was methodologically simpler and less expensive, making it a promising candidate for evaluation by national TB control programmes in developing countries.

Development of a clinical scoring system for the diagnosis of smear-negative pulmonary tuberculosis.

This study developed a clinical score based on clinical and radiographic data for the diagnosis of smear-negative pulmonary tuberculosis (SNPT). SNPT was defined as a positive culture in Ogawa in a patient with two negative sputum smears. Data from patients admitted to the emergency ward with respiratory symptoms and negative acid-fast bacilli (AFB) smears was analyzed by means of logistic regression to develop the predictive score.Two hundred and sixty two patients were included. Twenty patients had SNPT. The variables included in the final model were hemoptysis, weight loss, age > 45 years old, productive cough, upper-lobe infiltrate, and miliary infiltrate. With those, a score was constructed. The score values ranged from -2 to 6. The area under the curve for the ROC curve was 0.83 (95% CI 0.74-0.90). A score of value 0 or less was associated with a sensitivity of 93% and a score of more than 4 points was associated with a specificity of 92% for SNPT. Fifty-two point twenty-nine percent of patients had scores of less than one or more than four, what provided strong evidence against and in favor, respectively, for the diagnosis of SNPT. The score developed is a cheap and useful clinical tool for the diagnosis of SNPT and can be used to help therapeutic decisions in patients with suspicion of having SNPT.

Evaluation of the performance of two liquid-phase culture media for the diagnosis of pulmonary tuberculosis in a national hospital in Lima, Peru

OBJECTIVE To evaluate the diagnostic performance of two liquid-phase culture media for the diagnosis of pulmonary tuberculosis. PATIENTS AND METHODS From May to July 2003, sputum samples for culture were obtained from patients with respiratory symptoms attending the Hospital Nacional Cayetano Heredia. These were cultured in Ogawa medium, mycobacteria growth indicator tube (MGIT), and modified Middlebrook 7H9. Results were compared against a composite reference standard. RESULTS One hundred sputum specimens from 100 patients were included. Of these, 33 had culture-proven tuberculosis. The sensitivity of MGIT was found to be 100%. The modified Middlebrook 7H9 medium was found to have a sensitivity of 72.73%, while the sensitivity of Ogawa medium was found to be 69.70%. The mean growing time for MGIT was 12.18 days (95% confidence interval 10.24 to 14.12; p<0.01 vs. Ogawa and modified Middlebrook 7H9); for modified Middlebrook 7H9 was 16.65 days (95% confidence interval 14.85 to 18.80; p<0.01 vs. Ogawa), and for the Ogawa medium 25.74 days (95% confidence interval 22.22 to 29.6). CONCLUSIONS The liquid culture medium MGIT was superior to the modified Middlebrook 7H9 and the Ogawa media, both in terms of sensitivity and shorter growing time of colonies of Mycobacterium tuberculosis. The modified Middlebrook 7H9 medium is significantly faster but comparable in diagnostic performance to Ogawa. Costs remain an issue for MGIT.

Algorithm for the diagnosis of smear-negative pulmonary tuberculosis in high-incidence resource-constrained settings

Diagnosis of smear‐negative pulmonary tuberculosis (SNPT) remains a challenge, particularly in resource‐constrained settings. We evaluated a diagnostic algorithm that combines affordable laboratory tools and a clinical prediction rule (CPR).

Manual liquid culture on simple Middlebrook 7H9 or MGIT for the diagnosis of smear-negative pulmonary tuberculosis

To compare the performance of liquid culture on simple Middlebrook 7H9 to the one of manual mycobacterial growth indicator tube (MGIT) and solid culture on Ogawa for the diagnosis of smear‐negative tuberculosis (SN‐TB) in a high‐burden, resource‐constrained setting.

Detección de tuberculosis en el servicio de emergencia: utilidad de la segunda baciloscopía realizada el mismo día

Conducting a same day smears is a strategy that has recently been adopted by the World Health Organization. Our study sought to determine the additional diagnostic yield of a second sputum smear taken on the same day among patients with respiratory symptoms, attended at the emergency room from two hospitals in Lima. We included 270 patients from January to November 2011, which were asked two samples of sputum within two hours; the samples were processed by the Ziehl Neelsen method. The frequency of positive sputum smears was 18.5%. The additional diagnostic yield for the second smear was 20.9%. Considering the high prevalence of tuberculosis diagnosed by sputum smear, it is suggested that this test be included routinely in the emergency rooms of hospitals in Lima Metro area.