Displaying publications 21 - 40 of 54 in total

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  1. Sasidharan S, Lachumy SJ, Ravichandran M, Latha LY, Gegu SR
    Asian Pac J Trop Med, 2011 Jan;4(1):72-5.
    PMID: 21771421 DOI: 10.1016/S1995-7645(11)60037-0
    OBJECTIVE: To study the epidemiology of Helicobacter pylori (H. pylori) infection according to age group.

    METHODS: H. pylori infection data among 1 965 consecutive patients referred to the Endoscopy Unit collected at Sungai Petani Hospital for oesophagogastro-duodenoscopy (OGD). The patients were divided into 9 age groups (10-19, 20-29, 30-39, 40-49, 50-59, 60-69, 70-79, 80-89 and 90-99 years). In addition these groups were further divided into three minor group namely young adults (10-39), older adults (40-69) and geriatric groups (70-99).

    RESULTS: Overall prevalence of infection of H. pylori was analyzed and found that the prevalence increase with age (P<0.05). When the patients divided by ethnic and gender group with age, prevalence rate among young adults and older adults significantly higher (P<0.05) compared to geriatric groups across all races and gender (P<0.05). Furthermore, significantly higher number of males were infected compared to female (P<0.05) but such trend was only observed among older adult groups. In addition, there is a significant differences in H. pylori infection prevalence rates among ethnic groups (highest in Indians adults, followed Chinese and low in Malays, P<0.05).

    CONCLUSIONS: The overall prevalence of H. pylori did increase with age group across ethnicity and gender, in Northern Peninsular Malaysia.

  2. Chan M, Cheong TG, Kurunathan S, Chandrika M, Ledon T, Fando R, et al.
    Microb Pathog, 2010 Nov;49(5):211-6.
    PMID: 20558271 DOI: 10.1016/j.micpath.2010.06.001
    Cholera caused by the O139 serogroup still remains a public health concern in certain regions of the world and the existing O1 vaccines do not cross-protect cholera caused by this serogroup. An aminolevulinic acid (ALA) auxotroph vaccine candidate against the O139 serogroup, designated as VCUSM2, was recently developed. It was found to be immunogenic in animal model studies but showed mild reactogenic effects due to the presence of two intact copies of Vibrio cholerae toxin (CTX) genetic element. In the present study we have modified the ctx operon by systematic allelic replacement methodology to produce a mutant strain, designated as VCUSM14. This strain has two copies of chromosomally integrated and mutated ctxA gene, encoding immunogenic but not toxic cholera toxin A subunit (CT-A). The amino acids arginine and glutamic acid at position 7th and 112th, respectively, in CT-A of VCUSM14 were substituted with lysine (R7K) and glutamine (E112Q), respectively. Two copies of the ace and zot genes present in the ctx operon were also deleted. Cholera toxin-ELISA using GM1 ganglioside showed that the both wild type CT and mutated CT were recognized by anti-CT polyclonal antibodies. VCUSM14 produced comparatively less amount of antigenic cholera toxin when compared to the VCUSM2 and Bengal wild type strain. VCUSM14 did not elicit fluid accumulation when inoculated into rabbit ileal loops at doses of 10(6) and 10(8) CFU. The colonization efficiency of VCUSM14 was one log lower than the parent strain, VCUSM2, which can be attributed to the ALA auxotrophy and less invasive properties of VCUSM14. VCUSM14, thus a non-reactogenic auxotrophic vaccine candidate against infection by O139 V. cholerae.
  3. Asmal HS, Mustafa M, Abdullah S, Zaidah AR, Nurhaslindawati AR, Sarimah A, et al.
    PMID: 20578464
    Pneumocystis pneumonia (PCP) has become the most common opportunistic infection in HIV/AIDS patients with a CD4 count < or = 200. The incidence of PCP has declined as a result of prophylaxis and better highly active antiretroviral therapy (HAART). The objective of this study was to review the demographic data of HIV patients diagnosed clinically as having PCP at the Hospital Raja Perempuan Zainab II (HRPZ II) in Malaysia. This was a prospective study. All HIV patients admitted to HRPZ II with respiratory symptoms were enrolled in this study after giving informed consent. Their demographic data were collected. The total number of HIV patients reviewed in this study was 107. Nearly 60% of patients were clinically diagnosed as having pneumocystis pneumonia based on their signs, symptoms and chest x-ray findings. A CD4 count was available in 83 out of 107 patients. The fifty-three percent of patients(44) had a CD4 < 200 and were clinically diagnosed as having pneumocystis pneumonia. Thirty percent had a CD4 < 200 but did not have clinical pneumocystis pneumonia. Sixteen point nine percent had a CD4 > 200 and had clinical pneumocystis pneumonia, three of whom had received HAART, four patients had received prophylaxis. Overall, 94 patients (87.8%) received prophylaxis for pneumocystis pneumonia. Thirty-three patients (30.8%) received HAART. The occurrence of pneumocystis pneumonia was common before full implementation of HAART. Pneumocystis pneumonia can occur in patients with a CD4 >200.
  4. Al-Talib H, Yean CY, Al-khateeb A, Singh KK, Hasan H, Al-Jashamy K, et al.
    Curr Microbiol, 2010 Jul;61(1):1-6.
    PMID: 20033170 DOI: 10.1007/s00284-009-9567-8
    The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is responsible for nosocomial and community-acquired infections. Hence, rapid and accurate laboratory diagnosis of MRSA is a vital constituent of control measures. The present study evaluated five different methods for the identification of MRSA. A total of 207 S. aureus clinical isolates that consisted of 89 MRSA and 118 methicillin-susceptible S. aureus (MSSA) strains confirmed by PCR were tested. MRSA strains were evaluated by five different methods: chromogenic MRSA agar (CMRSA), oxacillin resistance screening agar base (ORSAB), mannitol salt oxacillin agar (MSO), mannitol salt cefoxitin agar with two different concentrations of cefoxitin [4 microg/ml (MSC-4) and 6 microg/ml (MSC-6)]. The results of the different methods were compared to mecA PCR as the gold standard. MSC-6 showed only six false-positive MRSA in comparison with PCR. The sensitivities and specificities of MSC-6, MSC-4, MSO-4, ORSAB, and CMRSA were as follows: 98.9/94.9%, 100/83.1%, 89.9/87.3%, 97.8/96.6%, and 95.5/94.9%, respectively. In comparison with PCR, it was found that both MSC-6 and ORSAB were relatively the least expensive screening tests ($0.70 and $1.00, respectively). In conclusion, all methods were comparable, but MSC-6 was the least expensive medium for MRSA screening.
  5. Cheong TG, Chan M, Kurunathan S, Ali SA, ZiNing T, Zainuddin ZF, et al.
    Microb Pathog, 2010 Feb;48(2):85-90.
    PMID: 19900531 DOI: 10.1016/j.micpath.2009.11.001
    Vibrio cholerae is a Gram-negative bacterium that causes diarrheal disease. V. cholerae O1 and O139 serogroups are toxigenic and are known to cause epidemic cholera. These serogroups produce cholera toxin and other accessory toxins such as accessory cholera enterotoxin, zonula occludens toxin, and multifunctional, autoprocessing repeat in toxin (MARTX). In the present study, we incorporated mutated rtxA and rtxC genes that encode MARTX toxin into the existing aminolevulinic acid (ALA) auxotrophic vaccine candidate VCUSM2 of V. cholerae O139 serogroup. The rtxC mutant was named VCUSM9 and the rtxC/rtxA mutant was named VCUSM10. VCUSM9 and VCUSM10 were able to colonize intestinal cells well, compared with the parent vaccine strain, and produced no fluid accumulation in a rabbit ileal loop model. Cell rounding and western blotting assays indicated that mutation of the rtxC gene alone (VCUSM9 strain) did not abolish MARTX toxicity. However mutation of both the rtxA and rtxC genes (VCUSM10) completely abolished MARTX toxicity. Thus we have produced a new, less reactogenic, auxotrophic rtxC/rtxA mutated vaccine candidate against O139 V. cholerae.
  6. Zaidah AR, Chan YY, Asma HS, Abdullah S, Nurhaslindawati AR, Salleh M, et al.
    PMID: 18564692
    This cross-sectional study determined the prevalence of cryptosporidiosis in HIV-infected patients using polymerase chain reaction (PCR). Stool specimens were collected from HIV infected patients who were admitted to Hospital Raja Perempuan Zainab II, Kota Bharu, Malaysia, for various indications from December 2004 to December 2005. A modified acid-fast stain was performed on the direct stool smears, then the stool specimens were further tested using nested PCR targeting the 18S rRNA gene of Cryptosporidium parvum, with a built-in internal control (IC). Out of 59 samples, 11 were positives. Nested PCR identified a total of nine samples (16%) compared to microscopy, which identified only three samples. All PCR negative results showed IC amplicons, suggesting that these samples were true negatives and were not due to inhibition of PCR. This study highlights the importance of molecular diagnosis in determining the true prevalence and epidemiology of C. parvum.
  7. Shareef BT, Harun A, Roziawati Y, Bahari IS, Deris ZZ, Ravichandran M
    J Contemp Dent Pract, 2008;9(3):114-20.
    PMID: 18335127
    This case report aims at describing an infection of the tongue as a manifestation of a Trichosporon asahii infection, its association with bronchial asthma and steroid administration, and to present a review of the literature pertaining to its antifungal susceptibility profile.
  8. Yean CY, Kamarudin B, Ozkan DA, Yin LS, Lalitha P, Ismail A, et al.
    Anal Chem, 2008 Apr 15;80(8):2774-9.
    PMID: 18311943 DOI: 10.1021/ac702333x
    A general purpose enzyme-based amperometric electrochemical genosensor assay was developed wherein polymerase chain reaction (PCR) amplicons labeled with both biotin and fluorescein were detected with peroxidase-conjugated antifluorescein antibody on a screen-printed carbon electrode (SPCE). As a proof of principle, the response selectivity of the genosensor was evaluated using PCR amplicons derived from lolB gene of Vibrio cholerae. Factors affecting immobilization, hybridization, and nonspecific binding were optimized to maximize sensitivity and reduce assay time. On the basis of the background amperometry signals obtained from nonspecific organisms and positive signals obtained from known V. cholerae, a threshold point of 4.20 microA signal was determined as positive. Under the optimum conditions, the limit of detection (LOD) of the assay was 10 CFU/mL of V. cholerae. The overall precision of this assay was good, with the coefficient of variation (CV) being 3.7% using SPCE and intermittent pulse amperometry (IPA) as an electrochemical technique. The assay is sensitive, safe, and cost-effective when compared to conventional agarose gel electrophoresis, real-time PCR, and other enzyme-linked assays for the detection of PCR amplicons. Furthermore, the use of a hand-held portable reader makes it suitable for use in the field.
  9. Gopinath VK, Al-Salihi KA, Yean CY, Ann MC, Ravichandran M
    J Clin Pediatr Dent, 2004;28(4):319-22.
    PMID: 15366620
    Amelogenesis imperfecta (AI) is a hereditary disorder resulting in generalized defects in the enamel. The case reported here is of a seven-year-old male child with yellow color of all his teeth. Two of his primary molars were extracted due to dental abscess with advanced root resorption. Histologically hypoplastic enamel layer, positively birefringent, generalized pitting, roughness with irregular general cracked borders were observed. Scanning electron microscope, revealed extensive irregular, disorganized rough superficial enamel layer. The enamel was irregularly decussate with filamentous prisms accompanied by small rounded formations. The morphological and histological examination of the tooth revealed that this patient has the features of AI. For genetic study blood sample were collected from the patient and PCR analysis revealed that there is no mutation in exons 1-7 of AMELX gene on the X chromosome of the patient. Hence, it is probable that the AI of this patient is not X-linked. It is more likely to be an autosomal mutation.
  10. Perumal V, Hashim U, Gopinath SC, Haarindraprasad R, Poopalan P, Liu WW, et al.
    Biosens Bioelectron, 2016 Apr 15;78:14-22.
    PMID: 26584078 DOI: 10.1016/j.bios.2015.10.083
    Creating novel nanostructures is a primary step for high-performance analytical sensing. Herein, a new worm like nanostructure with Zinc Oxide-gold (ZnO/Au) hybrid was fabricated through an aqueous hydrothermal method, by doping Au-nanoparticle (AuNP) on the growing ZnO lattice. During ZnO growth, fine tuning the solution temperature expedites random curving of ZnO nanorods and forms nano-worms. The nano-worms which were evidenced by morphological, physical and structural analyses, revealed elongated structures protruding from the surface (length: 1 µm; diameter: ~100 nm). The appropriate peaks for the face centred cubic gold were (111) and (200), as seen from X-ray diffractogram. The strong interrelation between Au and ZnO was manifested by X-ray photoelectron spectroscopy. The combined surface area increment from the nanoparticle radii and ZnO nanorod random curving gives raise an enhancement in detection sensitivity by increasing bio-loading. 'Au-decorated hybrid nano-worm' was immobilized with a probe DNA from Vibrio Cholera and duplexed with a target which was revealed by Fourier Transform Infrared Spectroscopy. Our novel Au-decorated hybrid nano-worm is suitable for high-performance bio-sensing, as evidenced by impedance spectroscopy, having higher-specificity and attained femtomolar (10 fM) sensitivity. Further, higher stability, reproducibility and regeneration on this sensing surface were demonstrated.
  11. Al-Talib H, Hasan H, Yean CY, Al-Ashwal SM, Ravichandran M
    Jpn J Infect Dis, 2011;64(1):58-60.
    PMID: 21266757
    Panton-Valentine leukocidin (PVL) is a cytotoxin which causes leukocyte destruction and tissue necrosis. Although it is produced by fewer than 5% of Staphylococcus aureus strains, PVL-producing S. aureus is emerging as a serious problem worldwide. There has been a marked increase in the incidence of necrotizing lung infections with a very high mortality associated with these strains. This report describes a fatal case of hospital-acquired necrotizing pneumonia caused by PVL-positive methicillin-susceptible S. aureus in a patient with a brain tumor.
  12. Al-Talib H, Yean CY, Al-Khateeb A, Hassan H, Singh KK, Al-Jashamy K, et al.
    BMC Microbiol, 2009;9:113.
    PMID: 19476638 DOI: 10.1186/1471-2180-9-113
    Staphylococcus aureus is a major human pathogen, especially methicillin-resistant S. aureus (MRSA), which causes a wide range of hospital and community-acquired infections worldwide. Conventional testing for detection of MRSA takes 2-5 days to yield complete information of the organism and its antibiotic sensitivity pattern.
  13. Meignanamoorthy M, Ravichandran M, Mohanavel V, Afzal A, Sathish T, Alamri S, et al.
    Materials (Basel), 2021 Aug 02;14(15).
    PMID: 34361508 DOI: 10.3390/ma14154315
    In this paper, Al-Fe-Si-Zn-Cu (AA8079) matrix composites with several weight percentages of B4C (0, 5, 10, and 15) were synthesized by powder metallurgy (PM). The essential amount of powders was milled to yield different compositions such as AA8079, AA8079-5 wt.%B4C, AA8079-10 wt.%B4C, and AA8079-15 wt.%B4C. The influence of powder metallurgy parameters on properties' density, hardness, and compressive strength was examined. The green compacts were produced at three various pressures: 300 MPa, 400 MPa, and 500 MPa. The fabricated green compacts were sintered at 375 °C, 475 °C, and 575 °C for the time period of 1, 2 and 3 h, respectively. Furthermore, the sintered samples were subjected to X-ray diffraction (XRD) analysis, Energy Dispersive Analysis (EDAX), and Scanning Electron Microscope (SEM) examinations. The SEM examination confirmed the uniform dispersal of B4C reinforcement with AA8079 matrix. Corrosion behavior of the composites samples was explored. From the studies, it is witnessed that the rise in PM process parameters enhances the density, hardness, compressive strength, and corrosion resistance.
  14. Xian TH, Sinniah K, Yean CY, Krishnamoorthy V, Bahari MB, Ravichandran M, et al.
    BMC Immunol, 2020 05 25;21(1):29.
    PMID: 32450807 DOI: 10.1186/s12865-020-00360-1
    BACKGROUND: Cholera, an acute watery diarrhoeal disease caused by Vibrio cholerae serogroup O1 and O139 across the continents. Replacing the existing WHO licensed killed multiple-dose oral cholera vaccines that demand 'cold chain supply' at 2-8 °C with a live, single-dose and cold chain-free vaccine would relieve the significant bottlenecks and cost determinants in cholera vaccination campaigns. In this direction, a prototype cold chain-free live attenuated cholera vaccine formulation (LACV) was developed against the toxigenic wild-type (WT) V. cholerae O139 serogroup. LACV was found stable and retained its viability (5 × 106 CFU/mL), purity and potency at room temperature (25 °C ± 2 °C, and 60% ± 5% relative humidity) for 140 days in contrast to all the existing WHO licensed cold-chain supply (2-8 °C) dependent killed oral cholera vaccines.

    RESULTS: The LACV was evaluated for its colonization potential, reactogenicity, immunogenicity and protective efficacy in animal models after its storage at room temperature for 140 days. In suckling mice colonization assay, the LACV recorded the highest recovery of (7.2 × 107 CFU/mL) compared to those of unformulated VCUSM14P (5.6 × 107 CFU/mL) and the WT O139 strain (3.5 × 107 CFU/mL). The LACV showed no reactogenicity even at an inoculation dose of 104-106 CFU/mL in a rabbit ileal loop model. The rabbits vaccinated with the LACV or unformulated VCUSM14P survived a challenge with WT O139 and showed no signs of diarrhoea or death in the reversible intestinal tie adult rabbit diarrhoea (RITARD) model. Vaccinated rabbits recorded a 275-fold increase in anti-CT IgG and a 15-fold increase in anti-CT IgA antibodies compared to those of rabbits vaccinated with unformulated VCUSM14P. Vibriocidal antibodies were increased by 31-fold with the LACV and 14-fold with unformulated VCUSM14P.

    CONCLUSION: The vaccine formulation mimics a natural infection, is non-reactogenic and highly immunogenic in vivo and protects animals from lethal wild-type V. cholerae O139 challenge. The single dose LACV formulation was found to be stable at room temperature (25 ± 2 °C) for 140 days and it would result in significant cost savings during mass cholera vaccination campaigns.

  15. Chua AL, Elina HT, Lim BH, Yean CY, Ravichandran M, Lalitha P
    J Med Microbiol, 2011 Apr;60(Pt 4):481-485.
    PMID: 21183596 DOI: 10.1099/jmm.0.027433-0
    Vibrio cholerae has caused severe outbreaks of cholera worldwide with thousands of recorded deaths annually. Molecular diagnosis for cholera has become increasingly important for rapid detection of cholera as the conventional methods are time-consuming and labour intensive. However, traditional PCR tests still require cold-chain transportation and storage as well as trained personnel to perform, which makes them user-unfriendly. The aim of this study was to develop a thermostabilized triplex PCR test for cholera which is in a ready-to-use form and requires no cold chain. The PCR test specifically detects both toxigenic and non-toxigenic strains of V. cholerae based on the cholera toxin A (ctxA) and outer-membrane lipoprotein (lolB) genes. The thermostabilized triplex PCR also incorporates an internal amplification control that helps to check for PCR inhibitors in samples. PCR reagents and the specific primers were lyophilized into a pellet form in the presence of trehalose, which acts as an enzyme stabilizer. The triplex PCR was validated with 174 bacteria-spiked stool specimens and was found to be 100 % sensitive and specific. The stability of the thermostabilized PCR was evaluated using the Q10 method and it was found to be stable for approximately 7 months at 24 °C. The limit of detection of the thermostabilized triplex PCR assay was 2×10(4) c.f.u. at the bacterial cell level and 100 pg DNA at the genomic DNA level, comparable to conventional PCR methods. In conclusion, a rapid thermostabilized triplex PCR assay was developed for detecting toxigenic and non-toxigenic V. cholerae which requires minimal pipetting steps and is cold chain-free.
  16. Kong JC, Sa'ad MA, Vijayan HM, Ravichandran M, Balakrishnan V, Tham SK, et al.
    Front Immunol, 2024;15:1384039.
    PMID: 38726000 DOI: 10.3389/fimmu.2024.1384039
    Chimeric antigen receptor-natural killer (CAR-NK) cell therapy is a novel immunotherapy targeting cancer cells via the generation of chimeric antigen receptors on NK cells which recognize specific cancer antigens. CAR-NK cell therapy is gaining attention nowadays owing to the ability of CAR-NK cells to release potent cytotoxicity against cancer cells without side effects such as cytokine release syndrome (CRS), neurotoxicity and graft-versus-host disease (GvHD). CAR-NK cells do not require antigen priming, thus enabling them to be used as "off-the-shelf" therapy. Nonetheless, CAR-NK cell therapy still possesses several challenges in eliminating cancer cells which reside in hypoxic and immunosuppressive tumor microenvironment. Therefore, this review is envisioned to explore the current advancements and limitations of CAR-NK cell therapy as well as discuss strategies to overcome the challenges faced by CAR-NK cell therapy. This review also aims to dissect the current status of clinical trials on CAR-NK cells and future recommendations for improving the effectiveness and safety of CAR-NK cell therapy.
  17. Amin A, Ali A, Kurunathan S, Cheong TG, Al-Jashamy KA, Jaafar H, et al.
    Histol Histopathol, 2009 05;24(5):559-65.
    PMID: 19283664 DOI: 10.14670/HH-24.559
    Vibrio cholerae is the causative agent of the infectious disease, cholera. The bacteria adhere to the mucosal membrane and release cholera toxin, leading to watery diarrhea. There are >100 serovars of V. cholerae, but the O1 and O139 serovars are the main causative agents of cholera. The present study aimed to compare the severity of intestinal mucosal infection caused by O1 El Tor and O139 V. cholerae in a rabbit ileal loop model. The results showed that although the fluid accumulation was similar in the loops inoculated with O1 and O139 V. cholerae, the presence of blood was detected only in the loops inoculated with the O139 serovar. Serosal hemorrhage was confirmed by histopathological examination and the loops inoculated with O139 showed massive destruction of villi and loss of intestinal glands. The submucosa and muscularis mucosa of the ileum showed the presence of edema with congested blood vessels, while severe hemorrhage was seen in the muscularis propria layer. The loops inoculated with O1 El Tor showed only minimal damage, with intact intestinal villi and glands. Diffuse colonies of the O139 serovar were seen to have infiltrated deep into the submucosal layer of the intestine. Although the infection caused by the O1 serovar was focal and invasive, it was more superficial than that due to O139, and involved only the villi. These observations were confirmed by immunostaining with O1 and O139 V. cholerae-specific monoclonal antibodies. The peroxidase reaction demonstrated involvement of tissues down to the submucosal layer in O139 V. cholerae infection, while in O1 El Tor infection, the reaction was confined mainly to the villi, and was greatly reduced in the submucosal region. This is the first reported study to clearly demonstrate the histopathological differences between infections caused by the O139 Bengal and O1 El Tor pathogenic serovars of V. cholerae.
  18. Deris ZZ, Van Rostenberghe H, Habsah H, Noraida R, Tan GC, Chan YY, et al.
    Int J Infect Dis, 2010 Jan;14(1):e73-4.
    PMID: 19482535 DOI: 10.1016/j.ijid.2009.03.005
    We report the first case of a human Burkholderia tropica infection. The patient was a premature neonate who had necrotizing enterocolitis with bowel perforation requiring surgical intervention. The stoma care and difficulties in feeding were a chronic problem. At the age of almost 4 months he developed septicemia due to B. tropica. Three consecutive blood cultures grew this organism. The organism was cleared from the blood after a course of imipenem and resolution of post-operative ileus. Our case suggests that environmental and plant pathogens can cause human infection especially in those in an immunocompromised condition.
  19. Lalitha P, Siti Suraiya MN, Lim KL, Lee SY, Nur Haslindawaty AR, Chan YY, et al.
    J Microbiol Methods, 2008 Sep;75(1):142-4.
    PMID: 18579241 DOI: 10.1016/j.mimet.2008.05.001
    A PCR assay has been developed based on a lolB (hemM) gene, which was found to be highly conserved among the Vibrio cholerae species but non-conserved among the other enteric bacteria. The lolB PCR detected all O1, O139 and non-O1/non-O139 serogroup and biotypes of V. cholerae. The analytical specificity of this assay was 100% while the analytical sensitivity was 10 pg/microL and 10(3) CFU/mL at DNA and bacterial level respectively. The diagnostic sensitivity and specificity was 98.5% and 100% respectively.
  20. Embong Z, Wan Hitam WH, Yean CY, Rashid NH, Kamarudin B, Abidin SK, et al.
    BMC Ophthalmol, 2008;8:7.
    PMID: 18445283 DOI: 10.1186/1471-2415-8-7
    The sensitivity and specificity of 18S rRNA polymerase chain reaction (PCR) in the detection of fungal aetiology of microbial keratitis was determined in thirty patients with clinical diagnosis of microbial keratitis.
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