Antimicrobial Susceptibility Patterns of Bacterial Isolates from Blood Culture of Pediatric Patients with Suspected Sepsis at a Tertiary Care Hospital in Mymensingh, Bangladesh.

Sepsis is a serious, life-threatening condition, occurring when an infectious agent invades the body, resulting in systemic inflammatory response syndrome (SIRS). Neonates and children are among the most vulnerable population groups of developing sepsis because of their weak immune barrier. Despite major advances in prevention, diagnosis and treatment of bacterial infections, invasive infections followed by sepsis remain one of the leading causes of childhood mortality. The aim of this study was to identify bacterial agents and antimicrobial resistance patterns of aerobic bacteria among children suspected of having sepsis. This cross-sectional descriptive type of observational study was conducted in the Department of Microbiology, Mymensingh Medical College, Bangladesh from March 2021 to February 2022. Blood samples were collected from pediatric patients, suspected of having sepsis referred from inpatient facility of department of Neonatology and Pediatrics, Mymensingh Medical College Hospital (MMCH). Blood samples were inoculated into BacT/ALERT PF Plus bottles followed by sub-culture of positive samples in blood agar, MacConkey agar and chocolate agar plates. Isolated bacteria were identified by routine biochemical tests. Antimicrobial resistance pattern of all isolated bacteria was seen by disk diffusion method. MIC of vancomycin by agar dilution method was determined for isolated S. aureus and Coagulase negative Staphylococci (CoNS). The prevalence of pediatric sepsis was 31.82% with highest isolation rate 35.55% among neonates. The isolation rate of gram-positive bacteria was 62.50% where S. aureus was the most common isolate 32.15% followed by CoNS 30.36%. Out of 21 gram-negative bacteria, Pseudomonas spp. was the most frequent isolate 7(33.33%), all of which were resistant to cefuroxime, ceftriaxone and ceftazidime along with all klebsiella and Acinetobacter isolates. Out of 18 S. aureus isolates, 94.44%, 88.89% and 66.67% were resistant to Azithromycin, Penicillin-G and Ciprofloxacin respectively. The MIC of Vancomycin by agar dilution method was observed <2µg/ml against all isolated S. aureus and CoNS. All the Gram-positive isolates were sensitive to Linezolid and Vancomycin. Early detection of bacteria followed by antimicrobial susceptibility test can help by selection of appropriate antibiotic and prevent spread of infection.

Detection of Oncogenic Human Papillomavirus (HPV-16 and HPV-18) from Bacterial Vaginosis Positive Patient Attending at Tertiary Care Hospital in Mymensingh.

Cervical cancer is the fourth most common cancer in women in the world and is the second leading malignancy among Bangladeshi women. Persistent infection with high risk human papillomavirus (HPV) is an important cause of development of cervical intraepithelial neoplasia (CIN) followed by cancer. Bacterial vaginosis (BV), a common treatable vaginal infection which can disrupt the balanced vaginal ecosystem and its innate protective mechanisms against infection, can play an essential role in the acquisition and persistence of high risk human papillomavirus (HR-HPV) infection. This cross sectional study was conducted to detect the HR-HPV (HPV-16 and HPV-18) infection among bacterial vaginosis positive patient in the Department of Microbiology, Mymensingh Medical College (MMC), Bangladesh, from March 2018 to February 2019. A total of 300 endocervical swabs and high vaginal swabs were collected from the VIA (Visual inspection with acetic acid) outdoor clinic of Obstetrics and Gynaecology Department of Mymensingh Medical college Hospital. HPV DNA was tested among all 300 cases by nested PCR. Typing of HPV 16 and HPV 18 was done among HPV DNA positive cases with BV and intermediate flora by multiplex PCR. BV was diagnosed according to Nugent criteria by using the gram stained smear of high vaginal swab. A total of 57/300 (19.0%) samples were positive for HPV DNA by nested PCR. Of the total 300 cases 78(26.0%) had BV, 38(13.0%) had intermediate flora and 184(61.0%) had normal vaginal flora. HPV DNA was more positive in patients having intermediate flora 08/38 (21.05%) followed by the patients having normal vaginal flora 37/184 (20.11%) and BV 12/78 (15.38%). Among the 12 BV patients who were also HPV DNA positive (83.33%) were belong to high risk HPV (type 16 and 18) group and among them 08(66.67%) were HPV-16 and 02(16.67%) were HPV-18. But among 08 HPV DNA positive intermediate flora containing patients only 01(12.5%) were belong to HR-HPV (type 16 and no type 18 was detected).

Species Identification and Antifungal Susceptibility Pattern of Candida Isolates in Patients with Vulvovaginitis from Mymensingh, Bangladesh.

Vulvovaginal Candidiasis (VVC), a frequent and cumbersome reproductive tract infection affects women's physical and mental health. Although Candida albicans was reported as the most common agent of VVC yet, recently there are significant changes in the pattern of Candida species causing VVC with varying antifungal susceptibility pattern. Therefore this cross-sectional, descriptive type of observational study conducted to identify the spectrum of Candida species associated with VVC and assesses their antifungal susceptibility pattern from March 2021 to February 2022. High vaginal swabs from 175 patients clinically suspected of VVC were collected and cultured on Sabouraud dextrose agar with Chloramphenicol. Species were identified by phenotypic methods like- germ tube test, sub-culture in chromogenic agar media and genotypic methods like- Polymerase chain reaction (PCR), Restriction fragment length polymorphism (RFLP). Antifungal susceptibility was done by disk diffusion method. Out of 175 patients, 52(29.7%) were positive for Candida species. Of the isolates- C. albicans 34(65.0%), Non albicans Candida (NAC) 18(35.0%). Among NAC, C. glabrata 5(9.6%), C. tropicalis 5(9.6%), C. parapsilosis 4(7.7%) and each of C. krusei, C. kefyr, C. ciferrii, C. dubliniensis were 1(1.9%). On susceptibility testing highest resistance was to Clotrimazole 31.0% followed by Nystatin 13.0%, Itraconazole 12.0% and Fluconazole 10.0%. Resistance to azole was higher in NAC than in albicans. Of these patients, 16(31.0%) had history of recurrent VVC (RVVC) of which 12(75.0%) were by NAC, predominantly C. glabrata 5(32.0%). The results showed the increasing incidence of NAC associated vaginitis with higher resistance and recurrence that should be considered in gynecology clinics.

Comparison of Wet Mount Microscopy and Giemsa Staining to PCR in the Diagnosis of Vaginal Trichomoniasis in a Tertiary Level Hospital of Bangladesh.

Trichomonas vaginalis (T vaginalis) is the most prevalent non-viral sexually transmitted infection of the reproductive age group, which may lead to various complications, if left untreated. This study aimed to diagnose Trichomonas vaginalis infection by different diagnostic procedures and to evaluate the efficacy of different diagnostic procedures. This cross-sectional descriptive study was conducted among 102 women with vaginal discharge at the Department of Obstetrics & Gynecology at Mymensingh Medical College Hospital (MMCH) from July 2019 to December 2020. Three ectocervical swabs were collected from each patient. Saline wet mount microscopy, giemsa staining and PCR were performed for each patient. Data were collected using a structured questionnaire and analyzed using Excel 2007, statistical package for social sciences (SPSS) version 26.0. The PCR assay detected Trichomonas vaginalis positivity in 6(5.9%) of 102 patients, followed by Giemsa staining 4.9% and Wet mount examination 2.9%. Wet mount microscopy showed less sensitivity 33.33%, but high specificity 98.95%, 66.67% positive predictive value, 95.96% negative predictive value and accuracy 95.09%. The sensitivity, specificity, PPV, NPV and accuracy of Giemsa staining were 66.67%, 98.96%, 80.0%, 97.94% and 97.06% respectively. Statistical significance was observed when both WMM and Giemsa staining were compared to gold standard test PCR. In resource limited settings, a wet mount is a good option for diagnosis of T vaginalis infection as giemsa staining requires heavy T vaginalis infection to be positive. But wherever facilities are available, PCR should be performed.

Prevalence of Virulence Genes in Acinetobacter baumannii Isolated from Clinical Samples in Mymensingh Medical College Hospital, Bangladesh.

Acinetobacter baumannii is an opportunistic bacterial pathogen that is the most important cause of hospital-acquired infections. The objective of this study was to evaluate the predominance and determination of virulence encoding genes in A. baumannii isolates. During this cross-sectional study period from February 2019 to March 2020 of 380 clinical samples including endotracheal aspirates (70), wound swab or pus (175), urine (70) and blood (65) analysed in inpatients admitted to the hospital in different unit like ICU, Surgery and Burn unit of Mymensingh Medical College Hospital. Out of 380 studied samples, 130(34.21%) strains were yielded growth. Among 130 isolates, Acinetobacter spp. was 49(37.69%). Totally, 39(79.59%) were Acinetobacter baumannii which was detected by molecular technique PCR. Further more, the determination of virulence genes csgA and fimH detected by PCR. Among two studied virulence genes, csgA (38.46%) was the most prevalent virulent genes associated with disease severity and co-morbidity of the patient in A. baumannii infections.

Distribution and Pattern of Anti-Tubercular Drug Resistance in Patients with Pulmonary Tuberculosis in Mymensingh Region of Bangladesh.

Globally, the emergence of multidrug-resistant strains of Mycobacterium tuberculosis is an increasing problem that adversely affects patient care and public health. This cross sectional descriptive study was carried out in the Department of Microbiology, Mymensingh Medical College from January 2010 to December 2010 to isolate M. tuberculosis from smear-positive sputum samples by Lowenstein-Jensen (L-J) media and investigate the drug resistance pattern. Among 101 smear-positive cases 80(79.20%) yielded growth of Mycobacteria, 5(4.95%) were contaminated and 16(15.84%) showed no growth. Among 80 isolates 76(95.0%) were M. tuberculosis and the remaining 4(5.0%) were Non-tuberculous Mycobacteria (NTM). Out of 76 M. tuberculosis 27(35.52%) were resistant to at least one drug, 4(5.26%) to Isoniazid (INH), 1(1.32%) to Rifampicin (RMP), 8(10.53%) to Streptomycin (SM) and 0(0.0%) to Ethambutol (EMB) and multi-drug resistant tuberculosis (MDR-TB) was 9(11.84%). The present study creates the impression that fairly high rate of anti-tuberculosis drug resistance among the tuberculosis cases and also high MDR-TB (Resistant to both Rifampicin and Isoniazide). The emergence of MDR-TB poses significant trouble to TB control activities throughout the world. The complexity of MDR-TB operation makes it essential to produce new skills to design, plan, application and monitor interventions for the management of MDR-TB. More surveillance and immediate remedial interventions should be performed to combat the trouble of MDR-TB to the general population.

Study of Human Brucellosis among Patients with Pyrexia of Unknown Origin by Antibody Detection.

This study was performed to determine the seropositivity of human brucellosis among the patients suffering from pyrexia of unidentified origin. This cross-sectional study was performed at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from September 2018 to August 2019; among the patients of pyrexia of unknown origin visited inpatient and outpatient facility of department of Medicine and department of Paediatrics, Mymensingh Medical College Hospital (MMCH) in Mymensingh division of Bangladesh. A total of 400 serum samples were screened by Brucella-specific latex agglutination test to determine seropositivity. Seven percent (7.0%) (28/400) serum samples were found to be seropositive for brucellosis by detecting Brucella-specific antibody at a titer ≥1:160. Therefore, Brucella-specific latex agglutination test may be recommended as a screening test for human brucellosis in developing and underdeveloped countries.

Molecular Detection of MBL Encoding Genes in Acinetobacter baumannii strains Isolated from Various Samples at a Tertiary Care Hospital in Mymensingh.

MBL producing Acinetobacter baumannii is a major threat for therapeutic treatment of hospital acquired infections. The aim of this study was to determine the prevalence of metallo-ß-lactamase genes VIM, IMP & SIM genes amongst isolated A. baumannii. This cross sectional study conducted in the department of Microbiology Mymensingh Medical College from March 2019 to February 2020. 49 Acinetobacter spp. were isolated from different clinical samples including endotracheal aspirates, wound swab/pus, urine and blood. Among 380 clinical samples 130 organisms were isolated growth was 34.21%. Out of 130 isolated strains, 49(37.69%) were Acinetobacter spp identified by standard bacteriological method and resistance to different antibiotics was assessed with Kirby- Bauer Disc diffusion method. Among 49 Acinetobacter spp, 39(79.59%) were Acinetobacter baumannii which was identified by molecular method PCR directing OXA-51 like gene. Multiplex PCR was done to determine MBL genes existence VIM, IMP & SIM. Ceftriaxone (79.48%) showing higher resistance and colistin (12.82%) showing lower resistance. All the strains were sensitive to tigecycline. The distribution of MBLs genes such as VIM 20(51.28%), IMP 5(12.82%) and SIM 0 (0%). This study showed that high level of antibiotic resistance and VIM was the most prevalent MBL genes among A. baumannii highlighting the need for indigenous antibiotic usage plan & infection control measures to prevent the spread of these resistance genes.

Concurrent Infection of Orientia Tsutsugamushi with Rickettsia spp. Including Rickettsia felis in North Central Bangladesh.

Rickettsial diseases are one of the leading causes of treatable acute febrile illness in Asia pacific region. This cross-sectional descriptive study was conducted at Department of Microbiology, Mymensingh Medical College to diagnose scrub typhus by rapid Immunochromatographic Test (ICT) and Nested PCR followed by molecular identification of possible Rickettsial coinfection among suspected febrile patients in Mymensingh, Bangladesh from March 2019 to February 2020. Among the enrolled 402 patients, 89 samples (22.13%) were seropositive by Immunochromatographic Test (ICT) and 65 samples (16.16%) were positive for O. tsutsugamushi DNA by Nested PCR, targeting 47KDa gene. Therefore, 113/402 (28.10%) samples were positive for scrub typhus by PCR and/or ICT. All the scrub typhus positive samples were further subjected to Nested PCR targeting 17 KDa gene for identification of Rickettsial co-infection and 13/113 (11.50%) were documented as positive. Then 13 Rickettsial co-infected samples were undertaken to automate sequencing and all were genetically confirmed as Rickettsia felis. Findings of the study may help clinicians to expand their list of differential diagnoses for undifferentiated fever and detection of Rickettsial co-infection may guide them to prescribe effective antimicrobials.

Efficacy of Probiotics in Treatment of Acute Rotavirus and Non Rotavirus Watery Diarrhoea in Children Admitted in Mymensingh Medical College Hospital.

Probiotics are live microorganisms that, when administered in adequate amount helps in reducing the duration of diarrhoea. Objective of this double blind randomized controlled clinical trial was to assess the efficacy of probiotics in treatment of acute rotavirus & non rotavirus watery diarrhoea among children aged 6 months to 2 years admitted at diarrhoea corner of Paediatrics Department of Mymensingh Medical College Hospital from October 2017 to May 2019. It was a double blind randomized controlled clinical trial. Total 500 sample were divided into Group A=ORS, zinc plus placebo (n=250) and Group B=ORS, zinc plus probiotics (n=250). Both Group A and Group B consisted of children presented with rotavirus and non-rotavirus diarrhoea. Placebo or probiotics were given once daily for 5 days which was prepared and coded by department of Pharmacology. Stool specimens were taken to Microbiology Department of MMCH for rotavirus detection. Rotavirus was detected by Polyacrylamide gel electrophoresis (PAGE). Data was analyzed by computer using SPSS program version 23.0. A total of 500 children with acute watery diarrhoea were included. Among them 188 children were diagnosed as rotavirus positive. Among group A found 89 rotaviral and 161 non rotaviral diarrhoea patients. Among group B found 99 rotaviral and 151 non rotaviral diarrhoea patients. Baseline characteristics were similar in both groups. The duration of diarrhoea, hospital stay, and fever was significantly lesser in probiotics group when compared with control (p<0.001). But duration of vomiting did not reduce significantly in probiotics group. Frequency of stools reduced significantly in probiotics group.

Coexistence of ESBL and MBL-mediated resistance in Acinetobacter baumannii in a Tertiary Care Hospital, Bangladesh.

Antimicrobial resistance mediated by extended-spectrum beta-lactamases (ESBL), AmpC beta-lactamase and metallo-beta-lactamase (MBL) producing Acinetobacter species is an emerging problem worldwide. In this cross-sectional study total 341 specimens were collected over a period of one year from January 2017 to January 2018. Specimens were collected from ICU and Surgery unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Specimens were collected from ICU and Surgery Unit of Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Samples were processed for culture by standard conventional methods and susceptibility testing and determined by Kirby Bauer disc diffusion method. Antibiotic discs and their strength were according to the CLSI 2017 guideline. Molecular study was done to detect the species by OXA-51 gene and drug resistance genes (IMP, VIM, NDM, TEM, SHV, CTX, SPM, SIM and GIM). Species identification was done by OXA-51 gene which is intrinsic to Acinetobacter baumannii. Among the 46 isolates, 36(78.26%) were positive for Oxa-51 gene, 16(34.8%) for TEM gene, 9(19.6%) for VIM gene, 3(6.5%) for NDM gene and 1(2.2%) for IMP gene. This study gives an alarming sign towards high prevalence of cephalosporin and carbapenem resistance due to production of extended spectrum beta-lactamases and metallo-betalactamases, respectively. Early detection, proper antibiotic policies, and compliance towards infection control practices are the best defenses against these organisms.

Socio-demographic and Clinico-epidemiological Study of Scrub Typhus in A Tertiary Care Hospital of Mymensingh, Bangladesh.

Scrub typhus is one of the leading causes of undifferentiated treatable febrile illness in Asia pacific region. It is grossly under diagnosed in many tropical countries of South Asia including Bangladesh, due to wide range of non-specific clinical presentations, low index of suspicion among clinicians, limited awareness and lack of accurate diagnostic facilities. This cross sectional observational study was conducted at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2020 enrolling 113 diagnosed cases of scrub typhus by Immunochromatographic test (ICT) and / or Nested PCR to characterize the socio-demographic and clinico-epidemiological features of scrub typhus in Mymensingh area. Majority of the scrub typhus cases came from rural areas (63.83%) and there was a slight female predominance (52.21%). The young (32.74%) and the young-adult age group (28.31%) were mostly affected. Most of the scrub typhus cases were housewives (30.98%), followed by farmers (23.89%) and students (21.23%). All the enrolled cases presented with fever. Other findings were myalgia (76.10%), headache (56.63%), cough (30.97%), vomiting (12.38%) and Respiratory distress (9.73%). Typical eschar of scrub typhus was present only in 9(7.96%) cases and 4(3.53%) patients had rashes on their skin. Few cases (3.53%) had jaundice and 15.96% cases were anaemic. Oliguria (7.96%) and neck rigidity (1.76%) were also documented. Most of the Nested PCR positive scrub typhus cases were documented during late rainy season and beginning of winter months. Findings of the study may offer increased awareness about high burden of scrub typhus as well as heightened suspicion among clinicians for early diagnosis, timely treatment and prevention of complications.

Detection of Quinolone Resistance Pattern and Presence of qnr Genes in Human Salmonella Isolates at Mymensingh, Bangladesh.

Among the quinolones, fluoroquinolones are broad spectrum antimicrobial agents used for treating many clinical infections including Salmonellosis. Although high level of resistance to fluoroquinolones remains low in Salmonella but reduced susceptibility is increasing worldwide. Plasmid-mediated quinolone resistance (PMQR) of qnr type (qnrA, B and S) has been identified now a day in several enterobacterial species including Salmonella spp. This cross-sectional study was held at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2020. This study was conducted to determine the current quinolone resistance pattern and to detect the presence of qnrA, qnrB and qnrS genes among Salmonella isolates. Antimicrobial susceptibility test of 36 Salmonella isolates were done by disc diffusion method. MIC of ciprofloxacin was detected by agar dilution method. Then amplification with specific primers of qnrA, qnrB and qnrS genes were performed for all Salmonella isolates. The present study observed 80.5% resistance to nalidixic acid, 33.3% to ciprofloxacin and 19.4% to ofloxacin by disc diffusion method. qnr A gene was detected in 2(5.5%) isolates, where as qnrS was detected in 5 (13.8%) isolates. None of the isolates was positive for qnrB gene. All the qnrA positive isolates showed resistance to Ciprofloxacin (MIC=128µg/ml) and Ofloxacin. In conclusion, presence of qnr genes in the study isolates is alarming, because, rapid dissemination might occur due to conjugative plasmid mediated horizontal transfer.

Seropositivity of Human Brucellosis among Patients with Pyrexia of Unknown Origin on Both Risk and Non-Risk Group of Individuals and Molecular Detection by Real-time PCR.

Brucellosis is a zoonotic disease that is one of the important infectious causes of Pyrexia of Unknown Origin (PUO). The objective of the present study was to determine the seropositivity and molecular detection of human brucellosis among the patients with pyrexia of unknown origin on both risk and non-risk group of individuals in greater Mymensingh. A total of 400 blood samples were randomly collected from pyretic patients started from September 2018 to August 2019. Questionnaires were used to collect data on both risk and non-risk group of individuals. All samples were initially screened for anti-Brucella antibodies using the Brucella-specific latex agglutination test. For accurate investigation, seropositive as well as seronegative serum samples were tested by BCSP31 Brucella genus-specific TaqMan real-time PCR. Overall 32(8%) cases were positive out of 400 samples by Brucella-specific latex agglutination test and/or BCSP31 Brucella genus-specific real-time PCR. Brucella-specific latex agglutination test documented 7% (28/400) positivity for brucellosis. 22(5.5%) samples found Brucella genus-specific real-time PCR positive out of 400 samples. Most real-time PCR positive cases were found from sero-positive samples of risk group population (15/32). Sero-negative but real-time PCR positive cases also found only from risk group population (4/32). There were 10 seropositive cases where real-time PCR was negative. In addition to Brucella-specific latex agglutination test as a screening test, Brucella genus-specific real-time PCR was performed for confirmation and also to avoid unjustified costs, drug toxicity, and masking of other potentially dangerous diseases.

Detection of Biocide Resistance Genes (qacE and qacΔE1) in Pseudomonas spp Isolated from Patients with CSOM at Mymensingh Medical College Hospital, Bangladesh.

Biocides, including disinfectants and antiseptics, are used for a variety of topical and hard surface applications in health care facilities. Biocides play a significant role for preventing and controlling nosocomial infections. However, failures in the antimicrobial activities of biocides have been reported. The resistance mechanism to disinfectants is usually determined by genes which are related to resistance to quaternary ammonium compounds, namely, qacE, qacΔE1 that are found in Gram-negative bacteria. The aim of this study is to detect the prevalence of Biocides resistance genes, qacE and qacΔE1, in clinical isolates of Pseudomonas spp. It was carried out from March 2017 to July 2018 in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Samples were collected from Outpatient of ENT department, MMCH. In this study, 300 clinical samples of CSOM cases were tested by the PCR method. The present study shows detection of biocide resistance genes (qacE, qacΔE1) among 87 isolated Pseudomonas spp by uniplex PCR. Among 72 clinical isolates of Pseudomonas aeruginosa 67(93.05%) had the gene qacEΔ1 and 25(34.72%) had the gene qacE. In addition other 15 Pseudomonas spp 3(20%) isolates had the qacEΔ1 gene and 2(13.33%) isolates had the qacE gene. In this study there is a marked difference in detection of the qacEΔ1 gene between the MDR and non MDR P. aeruginosa isolates. The qacEΔ1 was identified in 50 of 54(92.59%) MDR isolates and 7 of 18(38.89%) non MDR strains respectively. While gene qacE was detect 25(46.29%) MDR isolates and did not show any qacEΔ1gene in non MDR isolates. This study shows that the genes, qacE, qacΔE1 are widespread among Pseudomonas aeruginosa, they are higher in MDR strains than non MDR strains.

Rapid Serologic and Molecular Diagnosis of Scrub Typhus among Suspected Febrile Patients Visiting Mymensingh Medical College Hospital.

Scrub typhus, caused by the bacterium- Orientia tsutsugamushi is one of the leading causes of undifferentiated treatable febrile illness in Asia pacific region. It is grossly under diagnosed in many tropical countries of South Asia including Bangladesh, due to wide range of non-specific clinical presentations, low index of suspicion among clinicians, limited awareness and lack of accurate diagnostic facilities. This cross-sectional descriptive study was conducted at Department of Microbiology, Mymensingh Medical College to diagnose scrub typhus by rapid Immunochromatographic test (ICT) as well as molecular detection of O. tsutsugamushi by Nested PCR and automated nucleotide sequencing among suspected febrile patients in Mymensingh, Bangladesh during 2019-20. Blood samples were collected from 402 febrile patients of suspected Rickettsial illness, referred from inpatient and outpatient departments of Medicine and Pediatrics, Mymensingh Medical College Hospital (MMCH). Among the enrolled 402 patients, 89 samples (22.13%) were seropositive by Immunochromatographic test (ICT) and 65 samples (16.16%) were positive for O. tsutsugamushi DNA by Nested PCR, targeting 47KDa gene. Therefore, 113/402 (28.10%) samples were positive for scrub typhus by PCR and/ or ICT. Highest number of patients was detected positive by nested PCR during the first 5-10 days of fever but only 2 cases were positive after 20 days. In case of ICT, highest positivity for only IgM (8.13%) and both antibodies (2.43%) were documented in first 5-10 days of fever, but IgG positivity was highest (41.66) in >20 days of fever. From 65 PCR positive samples, automated nucleotide sequencing was performed on 20 randomly selected samples and all were genetically confirmed to be O. tsutsugamushi.

Early and Rapid Detection of Typhoid Fever by Nested PCR in Blood.

Typhoid fever caused by Salmonella typhi is one of the major health problems in developing countries including Bangladesh. Still now blood culture is gold standard method for diagnosing typhoid fever, but this method is laborious, requires several days and detection rate is low. Failure of early laboratory diagnosis often leads to increased morbidity and mortality. This study was intended to apply a nested PCR in blood for early diagnosis of typhoid fever. In this cross sectional study blood samples were collected from 200 suspected typhoid fever patients attending Mymensingh Medical College Hospital, Bangladesh. Nested Polymerase Chain Reaction (n PCR) of flagellin gene was done in all the blood samples. At the same time all blood samples were subjected to culture by lytic centrifugation method. Culture positive isolates were identified as S. typhi by biochemical tests. Among the 200 blood samples, 57 (28.5%) were positive for S. typhi on nested PCR where as blood culture was positive for S. typhi in 16 (8%) samples. Among the 57 PCR positive samples, only 15 (26.3%) samples were culture positive for S. typhi and rest 42 (73.7%) were culture negative. So, in culture negative cases PCR can be used as a rapid diagnostic test for diagnosing typhoid fever. Considering time requirement, PCR takes one day, whereas blood culture takes 3 or more days to confirm diagnosis.

Prevalence of ESBL Encoding Genes in Acinetobacter baumannii Strains Isolated from Various Samples of a Tertiary Care Hospital in Mymensingh.

The aim of this study was to find the prevalence of ESBL genes among A. baumannii isolates. In this cross sectional study, 49 Acinetobacter spp. were isolated from various clinical samples from March 2019 to February 2020 conducted in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Clinical samples including endotracheal aspirates, wound swab/pus, urine and blood. A total of 380 samples were analyzed. Growth was obtained in 34.21% of the samples yielding 130 organisms. Out of 130 organisms, 49(37.69%) were Acinetobacter spp. Among 49 Acinetobacter spp, 39(79.59%) were Acinetobacter baumannii which was identified by PCR targeting OXA-51 like gene. Amplification of the ESBL encoding genes, namely CTX-M, TEM, SHV done by molecular technique PCR. The most antibacterial resistance was against ceftriaxone (79.48%) and lower resistance only showed in colistin (12.82%). All the isolates were sensitive to tigecycline. The distribution of ESBLs genes such as TEM 20(51.28%), CTX-M 16(41.02%) and SHV 0(0%). The high resistance to most of the antibiotics among the studied strains and also a high prevalence of TEM gene in A. baumannii strains found in our study gives alarming sign towards the treatment complexity of these strains.