Enumeration of CD4, CD8 lymphocyte and CD4/CD8 ratio in HIV Infected patients by direct immunofluorescence

\ CD4 lymphocyte (T helper cell) plays a major role in the regulation of the immune response.\  Decrease in number of CD4 lymphocyte, due to being infected and destroyed by human immunodeficiency virus (Hiv), is the key index for monitoring the stage of disease progression.\  The appropriate treatment thus can be given to the patients at the right time corresponding to the need of the patients.\  By direct immunofluorescence technique using monoclonal antibodies, CD4, CD8 lymphocyte and CD4/CD8 ratio were enumerated and calculated.\  In 37 normal healthy controls (male 20, female 17), aged 26  5 years, the percentage of CD4 and CD8 lymphocytes were 38  8 (range : 23-60) and 28  7 (17-42), the absolute number of CD4 and CD8 lymphocytes were 956  277 cell/mm3 (545 \– 1504 cell/mm3 ) and 701  252 (348-1416) respectively.\  The CD4/CD8 ratio was 1.42  0.34 (0.72-2.48)\ We have studied 61 HIV-infected cases by categorizing them into asymtomatic HIV infection, AIDS relaed complex (ARC), and the full-blown AIDS.\  The results were tabulated in table 2.\  The best prognostic index was the absolute number of CD4 lymphocyte which could also differentiate among the various stages of HIV-infected patients; asymptomatic HIV infection (623  347; range : 92-1350), AIDS related complex (ARC) (347  365; 35-1509) and the full blown AIDS (137  130; 9-472).Key words : CD4 lymphocyte, CD4/CD8 ratio, AIDS, Immunofluorescence

Development of Fungal Media for using in Laboratory Learning

Background : Because common fungal media used in laboratory learning are expensive, and the nature of fungi in easily growing and the plenty of agricultural products in our country, can make it be possible to perform the experiment to prove the hypothesis and develop the cheaper media.Objective : To develop  the new fungal media for laboratory learning.Methods : Cultivate molds in the media each prepared from sweet potato, pumpkin, soy bean, mung bean and red bean, then select the one that yields the best fungal growth to develop the media formular for assessment of fungal growth comparing to the standard media.Results : Four formulae of the new fungal media containing pumpkin sucrose agar (PKSA), pumpkin dextrose agar (PKDA), red bean-pumpkin sucrose agar (RPSA), red bean-pumpkin dextrose agar (RPDA), were developed and tested to cultivate 7 molds compared with those of potato dextrose agar (PDA). Four species showed better growth in the 4 formulae and 2 species showed better growth in PDA, while 1 species showed no difference among all media (p

Detection and Differentiation of Mycobacterium Species in Clinical Specimens by Real-Time Polymerase Chain Reaction

Mycobacterium tuberculosis complex and Mycobacterium avium complex are causative agents of tuberculosis which is a major cause of human morbidity and mortality. Rapid diagnosis and species differentiation of mycobactria in specimens are important to control the disease and use appropriate antimicrobial therapy. In this study, primers and probe were design based on 16S-23S rDNA spacer sequence by using LightCycler 2.0 software. One set of primer, Rank 9, was used to perform real-time PCR and conventional PCR for detection and differentiation of Mycobacterium spp. Sybr Green dye for signal detection, successfully amplified all extracted DNA of mycobacteria. Its amplicon can be visualized by gel electrophoresis with the corresponding length of 214 bp. The nonspecific products of about 300 bp, were found in conventional PCR. The result suggested that these primers are not appropriate for conventional PCR. A hybridization probe for strain differentiation and verification of the protocol in clinical specimens is needed to complete the protocol.