We have evaluated a new set of primers (TRC 4 ) in comparison with the IS6110 primers commonly used in PCR to detect tuberculous meningitis among children. The levels of concordance between the results of IS6110 PCR and TRC 4 PCR with cerebrospinal fluid specimens from patients with clinically confirmed tuberculous meningitis were 80 and 86%, respectively. Results with the two primer sets were concordant for 55 positive and 22 negative specimens (n ‫؍‬ 98). We conclude that the sensitivity of PCR can be increased by using both IS6110 and TRC 4 primers.Tuberculosis (TB) is the major cause of death worldwide and is due to a single pathogen (16). Childhood TB in general and TB meningitis in particular owe their existence to unsuspected, undiagnosed, or incompletely treated adults in the community. Among children, mortality due to TB occurs mainly due to the neural form of TB, namely, TB meningitis. About 3% of the pediatric admissions to hospitals in India are due to TB meningitis, with reported mortality ranging from 17 to 71% (2, 14). Prompt diagnosis is critical for initiating appropriate therapy and facilitating measures to prevent dissemination of this highly contagious disease. The prevalence of childhood TB meningitis remains largely underestimated because clinical manifestations are nonspecific in early stages of the disease and bacteriologic confirmation is available only for a small proportion of patients. Also, clinical diagnosis of childhood TB meningitis is difficult due to its varied clinical presentations. Further, routinely used tests employed for clinical diagnosis of TB are inadequate to detect extrapulmonary forms of TB like TB meningitis.PCR is currently the most sensitive and rapid method to detect extrapulmonary Mycobacterium tuberculosis (1,5,8,11,15). We used as a new target TRC 4 , which was cloned and characterized previously in our laboratory (10). TRC 4 is a conserved repetitive element with specificity for M. tuberculosis complex. The aim of this paper was to compare the efficiency of a PCR with a target chosen from this cloned fragment with that of a PCR with the widely used IS6110 sequence in detecting M. tuberculosis in cerebrospinal fluid (CSF) samples from children with meningitis.CSF specimens from children suffering from meningitis, aged to 12 years, were included in the study if at least four of the following seven indicators of disease were in evidence: first, the presence of clinical features, such as gradual loss of playful activity, irritability, clouding of consciousness, convulsions, neck stiffness, and cranial nerve or motor defects; second, a CSF lymphocyte count at admission greater than 10 ϫ 10 6 / liter; third, a CSF protein level at admission greater than 80 mg/dl; fourth, a CSF glucose/blood glucose ratio at admission of less than 0.5; fifth, contact with an intrafamilial adult positive for pulmonary TB; sixth, induration of 10 mm or more on tuberculin testing with 1 TU of purified protein derivative; and seventh, positive radiological features of primary complex in...

The infective, microscopic Strongyloides stercoralis larvae in contaminated soil can penetrate human skin with the help of excretory/secretory proteases. These proteases play a critical role in infection and transmigration of the parasite to the intestines. Strongylastacin is similar to astacin (from the digestive gland of the crayfish Astacus astacus), a multi‐domain protein with a signal peptide, a pro‐enzyme, a catalytic domain containing the zinc binding consensus astacin family signature sequence HEXXHXXGFXHEXXRXDR, and a second conserved zinc binding motif SIMHY at N‐ terminal region. An EGF‐1 like domain and a CUB domain are located at the COOH‐ terminal. In this study, the excretory/secretory Strongylastacin gene from S. stercoralis infective larval stage was cloned and expressed as a 45 kDa in Escherichia coli. Immunoblot analysis showed the presence of natural IgG antibodies against strongylastacin in six infected and six non‐endemic normal sera. These findings were confirmed in an ELISA of 32 S. stercoralis infected and 32 presumed normal human sera; all contained natural anti‐strongylastacin IgG antibodies. By contrast, IgE antibodies specific to strongylastacin were present in sera from individuals infected with S. stercoralis but not in uninfected control sera. Moreover, recombinant strongylastacin did not cross‐react with IgE antibodies either from patients infected with filaria or patients with tropical pulmonary eosinophilic (TPE) who had increased IgE antibodies. The present authors conclude that strongylastacin, an excretory/secretory antigen, elicits specific IgE antibodies in S. stercoralis infected humans. Non‐specific IgG antibodies to strongylastacin are present in both infected and normal humans. Further investigation is needed to understand the role of the host protective response against strongylastacin.