ABSTRACT
@#<p style="text-align: justify;"><strong>BACKGROUND:</strong> Rabies is an important zoonotic disease that needs to be eradicated worldwide. It is still prevalent in the Philippines, thus development of a relatively affordable but still accurate and rapid post-mortem detection test for the virus is desired, especially in regional laboratories.<br /><strong>METHODS:</strong>The study evaluated the Direct Rapid Immunohistochemical Testing (DRIT) of hippocampal touch impressions of suspected rabid Canis lupus familiaris using monospecific N protein polyclonal antibody developed by the Research Institute for Tropical Medicine (RITM). One hundred sixty (160) acetone-fixed hippocampal touch impressions were subjected DRIT.<br /><strong>RESULTS:</strong> One hundred thirteen (70.6%) out of 160 samples tested positive for rabies viral antigen (RVA) and 47 (29.4%) out of 160 samples tested negative for RVA. No false positive and false negative results were obtained. The results agree with the gold standard, dFAT.<br /><strong>CONCLUSION:</strong> DRIT was able to detect low to high concentrations of RVA in the hippocampal touch impressions based on the grading distribution. DRIT had 100% sensitivity, specificity and over-all accuracy using monospecific polyclonal antibodies, which suggests its use as a more affordable alternative to the gold standard dFAT.</p>
Subject(s)
Dogs , Animals , Antigens, Viral , Hippocampus , Rabies , Rabies virus , Sensitivity and Specificity , Touch , Tropical Medicine , ImmunohistochemistryABSTRACT
<p><b>AIM</b>To establish a sensitive and efficient reporter gene-based screening model for finding agonists of estrogen receptor beta subtype.</p><p><b>METHODS</b>A recombinant vector pTAL-ERE-SEAP was constructed by inserting a synthetic sequence composed of five estrogen responsive elements in front of promoter of pTAL-SEAP vector. pTAL-ERE-SEAP was then transfected into human embryonic kidney (HEK293) cells. G418 (200 microg x mL(-1)) was added to select positive clones that can be induced by E2 to express reporter gene SEAP. The speciality was tested by several ligands of relative nuclear receptors of the same family. The stability of the model, the time-effect relationship, the dose-response relationship, and the immunocytochemistry staining of ERbeta expression after transfection were observed. 2 622 compounds were screened by using this model.</p><p><b>RESULTS</b>Stably transfected clones were obtained. The expression levels of reporter gene SEAP of positive clones was induced by E2 in a dose-response and time-effect relationship manners. The Z' factor value was 0.7. The expression levels of dexamethasone and other ligands were low. The result of immunocytochemistry staining showed the expression of ERbeta. E2 had no proliferating effects on stably transfected clones.</p><p><b>CONCLUSION</b>Stably transfected positive clones transfected with recombinant vector pTAL-ERE-SEAP were obtained. The positive clones may be used to screen for agonists of estrogen receptor beta subtype by measurement of luminescent value of expressed SEAP in wells of microlitre plate.</p>