Patients' Readiness Towards Teledentistry in the Malaysian Urban Population Attending an Undergraduate Teaching University.

Introduction: COVID-19 disease has resulted in suspension of all nonurgent routine dental treatments. In view of COVID-19 situation, social distancing, movement restriction orders, and affected health care systems, there is an urgent need to resume and deliver oral health care remotely. Hence, alternative means of dental care should be available for both patients and dentists. Therefore, this study aims to assess patients' readiness for teledentistry in Malaysian urban population attending an undergraduate teaching university. Methods: A cross-sectional study was conducted among 631 adult patients visiting the Faculty of Dentistry, SEGi University, from January 2020 to May 2021 in Selangor, Malaysia. A validated, self-administered, 5-point Likert scale online questionnaire comprising five domains was administered. (1) Patients' demographics and dental history, (2) patients' access to teledentistry, (3) patients' understanding towards teledentistry, (4) patients' willingness, and (5) barriers in using teledentistry were used to collect the required information. Results: Six hundred and thirty-one (n = 631) participants responded to the questionnaire. Ninety percent of patients were able to connect to Wi-Fi services independently and 77% participants were comfortable using online communication platforms. Seventy-one percent of the participants agreed that video and telephone clinics can reduce chances of infection rather than face-to-face consultation during the pandemic. Fifty-five percent of patients felt that virtual clinics would save time and 60% thought it could reduce travelling costs. Fifty-one percent showed their willingness to use video or telephone clinics when implemented at onsite clinics. Conclusion: Our study shows the readiness of patients to accept teledentistry as an alternative method of oral care if appropriate training and education are provided. The results of this study have prompted an increase in patients' education and shown a need to train clinicians and patients to integrate this technology at SEGi University. This might facilitate unhindered dental consultation and care in all situations.

Humoral and T cell responses to SARS-CoV-2 reveal insights into immunity during the early pandemic period in Pakistan.

BACKGROUND: Protection against SARS-CoV-2 is mediated by humoral and T cell responses. Pakistan faced relatively low morbidity and mortality from COVID-19 through the pandemic. To examine the role of prior immunity in the population, we studied IgG antibody response levels, virus neutralizing activity and T cell reactivity to Spike protein in a healthy control group (HG) as compared with COVID-19 cases and individuals from the pre-pandemic period (PP). METHODS: HG and COVID-19 participants were recruited between October 2020 and May 2021. Pre-pandemic sera was collected before 2018. IgG antibodies against Spike and its Receptor Binding Domain (RBD) were determined by ELISA. Virus neutralization activity was determined using a PCR-based micro-neutralization assay. T cell - IFN-γ activation was assessed by ELISpot. RESULTS: Overall, the magnitude of anti-Spike IgG antibody levels as well as seropositivity was greatest in COVID-19 cases (90%) as compared with HG (39.8%) and PP (12.2%). During the study period, Pakistan experienced three COVID-19 waves. We observed that IgG seropositivity to Spike in HG increased from 10.3 to 83.5% during the study, whilst seropositivity to RBD increased from 7.5 to 33.3%. IgG antibodies to Spike and RBD were correlated positively in all three study groups. Virus neutralizing activity was identified in sera of COVID-19, HG and PP. Spike reactive T cells were present in COVID-19, HG and PP groups. Individuals with reactive T cells included those with and without IgG antibodies to Spike. CONCLUSIONS: Antibody and T cell responses to Spike protein in individuals from the pre-pandemic period suggest prior immunity against SARS-CoV-2, most likely from cross-reactive responses. The rising seroprevalence observed in healthy individuals through the pandemic without known COVID-19 may be due to the activation of adaptive immunity from cross-reactive memory B and T cells. This may explain the more favourable COVID-19 outcomes observed in this population.

Progressive multifocal leukoencephalopathy in an immunocompetent patient: A case report and review of literature.

Progressive multifocal leukoencephalopathy (PML), a demyelinating disease of the brain, caused by the John Cunningham virus (JCV) is usually seen in patients who are immunocompromised. Here, we describe a case of an immunocompetent patient diagnosed with PML and a comprehensive literature review. A 64-year-old Caucasian male presented with acute worsening of progressive neurological decline with difficulty in vision and reading. Based on history, examination, cerebrospinal fluid markers, histopathology, and magnetic resonance imaging brain at the time of presentation diagnosed the patient with PML in a setting of no immunosuppression disorder. The patient was started on Pelfilgrastim with significant systematic improvement. In our literature review, it was seen that the average age of symptom presentation was 57.5 with predominance in males. Most of the patients presented with progressive neurological deficits with symptomology ranging from mild confusion, aphasia, anxiety to sensory disturbances with numbness, hemiparesis, and hemianopsia. Out of the 21 cases, patients responded to mirtazapine and intravenous pulse methylprednisolone (IVMP). The mortality rate was close to 50% with 11 fatal cases and 10 nonfatal cases. Our case and literature review demonstrate the possibility that PML may very rarely occur in patients that are immunocompetent. Furthermore, our review showed that patients responded well to mirtazapine and IVMP. We also want to highlight that the mortality rate was lower in this review and was only compared to mortality in PML associated with immunocompromised status.

Effect of lip thickness and competency on soft-tissue changes.

INTRODUCTION: Improvement in facial esthetics is a major reason for seeking orthodontic treatment. Soft tissues responsible for esthetics show a variable response to the movement of underlying teeth during orthodontics. METHODS: The purpose of this cross-sectional study was to analyze the response of soft tissues and compare it among groups presenting with different lip thickness and competence. It was carried out on 37 patients with Class I and Class II Division 1 malocclusion who had undergone extractions of maxillary first premolars and had completed their orthodontic treatment. Data were obtained by corresponding lateral cephalometric radiographs taken before and at the end of orthodontic treatment. Association was analyzed with a Pearson correlation test. Differences among groups exhibiting varying competency and lip thickness were tested with an independent sample t test. A P value of ≤0.05 was considered statistically significant. RESULTS: Pearson correlation revealed significant associations among incisor tip retraction with lip base retraction (r = 0.68), lip base thinning (r = 0.41) and vermilion retraction (UV) (r = 0.73). Regression analysis showed a moderate increase in lip length (AB) and thickness at vermilion, which were 0.11 mm and 0.15 mm for each mm of incisor retraction at the tip, whereas stronger effects were observed for UV (0.38 mm) and lip base retraction (0.55 mm). There was significantly more lip base thinning (P = 0.03) and UV (P = 0.04) in the incompetent group compared with the competent group. The AB increased significantly in the sample with thicker lips (P = 0.01). Pearson correlation coefficient showed a strong association of lip retraction at the base and vermilion with the incisor movement at the cervical area, for competent and incompetent lips, and thicker and thinner lips. CONCLUSIONS: Lip retraction at vermilion and lip base thinning was significantly more in patients with incompetent lips, whereas the AB increased significantly more in the group with thicker lips.

Relationship of early weaning and non-nutritive sucking habits with facial development.

OBJECTIVE: To determine the frequency of malocclusion features, convex facial profile and non-nutritive sucking habits among children with and without early weaning. METHODS: The cross-sectional study was conducted at the National Institute of Child Health, Karachi, from June to December 2020, and comprised of either gender aged 3-6 years. The subjects were clinically examined, and details of lactation period, bottle feeding, thumb and pacifiers sucking were gathered from the parents to assess association with malocclusion. Data was analysed using SPSS 20. RESULTS: Of the 180 subjects, 100(55.5%) were boys and 80(45.5%) were girls. There was significant association of early weaning with non-nutritive sucking habits, absent primate spaces, deviated midlines, and complete overbite (p<0.05). Breastfeeding <2 years was significantly associated with anterior crossbite (p<0.05). Early weaning had no association with convex facial profile, distoclusion, anterior and posterior crossbite and increased overjet (p>0.05). Non-nutritive sucking habits had significant relationship with deviated dental midlines (p>0.05). Conclusion: Early weaning was found to be a factor in establishing non-nutritive sucking habits, and was strongly associated with crowding, disturbed vertical overlapping of incisors and developing decreased facial height. Breastfeeding for less than the recommended duration had a tendency to lead to anterior cross-bite.

COVID-19 and Mucormycosis Coinfection: How Challenging It Is.

Recently, cases of rhinoorbital mucormycosis in people diagnosed with COVID-19 have been reported from India particularly. Diabetes mellitus though happens to be an independent risk factor both for severe COVID-19 and mucormycosis, administration of steroids is attributed as a precipitating factor for acquiring the comorbid condition. This opportunistic fungal infection is highly angioinvasive in nature because of which, clinical outcome of infection is invariably poor, especially with rhinocerebral or rhinoorbitocerebral variety of mucormycosis. However, effective management depends upon timely and accurate diagnosis and parenteral administration of amphotericin B. At the same time, judicious use of steroids is a key factor. In addition, glycemic control in those who are severely diabetic is strongly advocated. Exenteration of an eyeball may be indicated if cavernous sinus and intracranial spread are anticipated. Therefore, in order to facilitate faster healing and better penetration of antifungal drugs, surgical debridement of the paranasal sinus cavities and removal of dead tissue from the sinuses are recommended.

Anti-depressants and COVID-19: A New Ray of Hope.

The coronavirus disease pandemic has grown worldwide. As we understand the exact pathophysiology of the disease and how it affects the systems in the human body, we are in the process of discovering and repositioning drugs potentially effective in these regards. A few targets of these drugs are excessive inflammation following SARS-CoV-2 infection and sigma-1 receptor ER chaperone protein, which plays a role in replication. The recent discovery of antidepressants like fluvoxamine and clomipramine acting through these targets may provide a new ray of hope to decrease mortality and morbidity in severe COVID patients.

Mucormycosis and COVID-19: An epidemic within a pandemic in India.

IMPORTANCE: Coronavirus disease (COVID-19) causes an immunosuppressed state and increases risk of secondary infections like mucormycosis. We evaluated clinical features, predisposing factors, diagnosis and outcomes for mucormycosis among patients with COVID-19 infection. METHODS: This prospective, observational, multi-centre study included 47 consecutive patients with mucormycosis, diagnosed during their course of COVID-19 illness, between January 3 and March 27, 2021. Data regarding demography, underlying medical conditions, COVID-19 illness and treatment were collected. Clinical presentations of mucormycosis, imaging and biochemical characteristics and outcome were recorded. RESULTS: Of the 2567 COVID-19 patients admitted to 3 tertiary centres, 47 (1.8%) were diagnosed with mucormycosis. Mean age was 55 ± 12.8years, and majority suffered from diabetes mellitus (n = 36, 76.6%). Most were not COVID-19 vaccinated (n = 31, 66.0%) and majority (n = 43, 91.5%) had developed moderate-to-severe pneumonia, while 20 (42.6%) required invasive ventilation. All patients had received corticosteroids and broad-spectrum antibiotics while most (n = 37, 78.7%) received at least one anti-viral medication. Mean time elapsed from COVID-19 diagnosis to mucormycosis was 12.1 ± 4.6days. Eleven (23.4%) subjects succumbed to their disease, mostly (n = 8, 72.7%) within 7 days of diagnosis. Among the patients who died, 10 (90.9%) had pre-existing diabetes mellitus, only 2 (18.2%) had received just one vaccine dose and all developed moderate-to-severe pneumonia, requiring oxygen supplementation and mechanical ventilation. CONCLUSIONS: Mucormycosis can occur among COVID-19 patients, especially with poor glycaemic control, widespread and injudicious use of corticosteroids and broad-spectrum antibiotics, and invasive ventilation. Owing to the high mortality, high index of suspicion is required to ensure timely diagnosis and appropriate treatment in high-risk populations.

A qualitative study of trainer and trainee perceptions and experiences of clinical assessment in post-graduate dental training.

BACKGROUND: The implementation of workplace-based assessment (WBA) needs to ensure the achievement of pre-set competences but may look different across varying contexts, such as in post-graduate dental education. The purpose of this study is to explore the perception of residents, faculty members and alumni concerning their experience with clinical assessment, and what configurations they consider as optimal to maximise the entailed learning experience. METHODS: This study relied on a qualitative descriptive design using two data collection tools: focus group sessions, and semi-structured, one-to-one interviews. Data were triangulated from three sources: residents, faculty members and alumni. The data were inductively analysed based on constructivist epistemology. This was done using the Thematic Analysis approach, facilitated by NVivo software. RESULTS: The analysis revealed two mutually exclusive themes: process and people. Within process, variables related to quality, workflow and feedback surfaced. As for the people theme, the main two group of stakeholders referred to in the related analysis were the trainees and the trainers. DISCUSSION: There are many variables that need to be considered when developing an evidence-driven WBA. In addition, factoring into the design of the WBA the perception of the main stakeholders will enable contextualisation which is expected to raise the reliability of the adapted tools. CONCLUSION: This study introduced a framework that could support post-graduate universities in their journey towards developing context-specific WBA.

Chikungunya outbreak in Karachi Pakistan 2016-2017: An analysis of viral isolates.

In December 2016 physicians in Karachi, Pakistan,witnessed an increase in patients presenting with febrile illness and severe polyarthralgia. Subsequently, chikungunya virus (CHIKV)) was isolated from three patients. This virus was sequenced and compared with other isolates of CHIKV obtained in India and Pakistan during recent outbreaks. Phylogenetic analysis indicated that the Karachi isolates were most similar to the East Central South African CHIKV lineage and showed sequence homology to isolates obtained in other parts of Pakistan and India. More importantly, two of the CHIKV isolates had a nucleotide substitution in the E1 gene corresponding to an amino acid change at chain F portion of the E1 protein.

Increase in Penicillin and multidrug resistance in Streptococcus pneumoniae (1993-2016): Report from a tertiary care hospital laboratory, Pakistan.

OBJECTIVE: To determine the trend of resistance to antimicrobials in Streptococcus pneumoniae infections, and the impact of new Clinical and Laboratory Standards Institute guidelines on 1211 among meningeal isolates. METHODS: The descriptive observational retrospective study was conducted at the Aga Khan University Hospital laboratory in Karachi, and comprised Streptococcus pneumoniae isolation and antimicrobial susceptibility data over a period of 24 years, from 1993 to 2016, which was compared in terms of pre-2008 and post-2008 data, which was analysed using SPSS 19. RESULTS: Of the 7415 non-duplicate isolates identified, 4700(63.4%) were from male patients and 2,715(36.6%) were from female patients. The overall mean age of the patients was 38±27 years. Penicillin resistance in non-meningeal isolates during the two periods was not significantly different (p>0.05), but a significant rise in penicillin resistance in meningeal isolates was observed in the second period (p<0.05). High resistance rates were observed for co-trimoxazole, tetracycline and erythromycin, and an increased trend of multi-drug resistant strains was also noted from 1999 {n=35/317(11%)} to 2016 {n=110/314 (36%)}. CONCLUSIONS: The emergence of multi-drug resistant strains was evident. The spike in penicillin-resistant Streptococcus pneumoniae in meningeal isolates may have been due to the revised guidelines by the Clinical and Laboratory Standards Institute.

Carbapenem-Resistant Acinetobacter baumannii and Enterobacteriaceae in South and Southeast Asia.

Carbapenem-resistant Gram-negative bacteria, in particular the Acinetobacter baumannii-calcoaceticus complex and Enterobacteriaceae, are escalating global public health threats. We review the epidemiology and prevalence of these carbapenem-resistant Gram-negative bacteria among countries in South and Southeast Asia, where the rates of resistance are some of the highest in the world. These countries house more than a third of the world's population, and several are also major medical tourism destinations. There are significant data gaps, and the almost universal lack of comprehensive surveillance programs that include molecular epidemiologic testing has made it difficult to understand the origins and extent of the problem in depth. A complex combination of factors such as inappropriate prescription of antibiotics, overstretched health systems, and international travel (including the phenomenon of medical tourism) probably led to the rapid rise and spread of these bacteria in hospitals in South and Southeast Asia. In India, Pakistan, and Vietnam, carbapenem-resistant Enterobacteriaceae have also been found in the environment and community, likely as a consequence of poor environmental hygiene and sanitation. Considerable political will and effort, including from countries outside these regions, are vital in order to reduce the prevalence of such bacteria in South and Southeast Asia and prevent their global spread.

Prognostic significance of epidermal growth factor receptor (EGFR) over expression in urothelial carcinoma of urinary bladder.

BACKGROUND: Epidermal growth factor receptor (EGFR) has been shown to have abnormal expression in many human cancers and is considered as a marker of poor prognosis. Frequency of over expression in bladder cancer has not been studied in our population; therefore we aimed to evaluate the frequency and prognostic significance of EGFR immunohistochemical expression in locoregional population. METHODS: We performed EGFR immunohistochemistry on 126 cases of bladder cancer and association of EGFR expression with tumor grade, lamina propria invasion, deep muscle invasion and recurrence of disease was evaluated. RESULTS: High EGFR expression was noted in 26.2% (33 cases), 15.1% (19 cases) and 58.7% (74 cases) revealed low and no EGFR expression respectively. Significant association of EGFR expression was noted with tumor grade, lamina propria invasion, deep muscle invasion and recurrence status while no significant association was seen with age, gender and overall survival. Kaplan- Meier curves revealed significant association of EGFR expression with recurrence while no significant association was seen with overall survival. CONCLUSION: Significant association of EGFR overexpression with tumor grade, muscularis propria invasion and recurrence signifies its prognostic value; therefore EGFR can be used as a prognostic biomarker in Urothelial bladder carcinoma.

Prognostic significance of p16 & p53 immunohistochemical expression in triple negative breast cancer.

Background: p16 and p53 genes are frequently mutated in triple negative breast cancer & prognostic value of these mutations have been shown; however, their role as immunohistochemical overexpression has not been fully validated. Therefore we aimed to evaluate the association of p16 and p53 overexpression in triple negative breast cancer with various prognostic parameters. Methods: Total 150 cases of triple negative breast cancers were selected from records of pathology department archives that underwent surgeries at Liaquat National hospital, Karachi from January 2008 till December 2013. ER, PR and Her2neu immunohistochemistry were re-performed to confirm triple negative status. p16 & p53 immunohistochemistry was performed on all cases and association with various clinicopathologic parameters was determined. Results: Mean age of the patients involved in the study was 48.9 years. Most of the patients presented at stage T2 with a high mean ki67 index i.e. 46.9%. 42.7% of cases had nodal metastasis. Although 84% cases were of invasive ductal carcinoma; however a significant proportion of cases were of metaplastic histology (9.3%). Fifty-one percent (76 cases) of cases showed positive p53 expression while 49% (74 cases) were negative. Higher percentage of p53 expression was found to correlate with higher T stage, high ki67 index and higher nodal stage. On the other hand, strong intensity of p53 expression was positively correlated with higher tumor grade and ki67 index. Seventy-one percent (98 cases) of cases showed positive p16 expression, whereas 24.8% (34 cases) were negative and 3.6% (5 cases) showed focal positive p16 expression. However, no significant association was found between p16 expression and various clinical and pathologic parameters. Similarly, no significant association of either p16 or p53 over-expression was noted with recurrence status of patients. Conclusion: On the basis of significant association of p53 over-expression with worse prognostic factors in triple negative breast cancer, therefore we suggest that more large scale studies are needed to validate this finding in loco-regional population. Moreover, high expression of p16 in triple negative breast cancer suggests a potential role of this biomarker in triple negative breast cancer pathogenesis which should be investigated with molecular based research in our population.

Immunohistochemical expression of epidermal growth factor receptor (EGFR) in South Asian head and neck squamous cell carcinoma: association with various risk factors and clinico-pathologic and prognostic parameters.

BACKGROUND: In this study, we intend to determine the immunohistochemical expression of EGFR in cases of head and neck squamous cell carcinoma and its association with prognostic clinico-pathologic features. METHODS: A total of 115 cases of head and neck squamous cell carcinoma treated at Liaquat National Hospital, Karachi, Pakistan, were included in the study. Clinico-pathologic features, risk factors, and recurrence status of cases were evaluated, and EGFR immunohistochemistry was performed. RESULTS: In our study, 52 cases (45.2%) of head and neck SCC were positive and 10 cases (8.7%) were focal positive for EGFR expression, while 53 cases (46.1%) were negative for EGFR expression. High EGFR expression (> 70%) was noted in 6.1% (7 cases), while 12.2% (14 cases) and 26.1% (30 cases) revealed 51-70% and 11-50% EGFR expression respectively. On the basis of intensity, strong EGFR expression was noted in 13.9% (16 cases) while 16.5% (19 cases) and 23.5% (27 cases) revealed intermediate and weak EGFR expression respectively. Significant association of EGFR expression was noted with tumor stage and disease-free survival. CONCLUSION: We found a significant association of EGFR expression with tumor stage and disease-free survivals, which are the most important prognostic factors in head and neck squamous cell carcinoma; therefore, EGFR expression can help as a prognostic biomarker in head and neck squamous cell carcinoma. On the other hand, we suggest that molecular studies should be performed in squamous cell carcinoma of head and neck in our setup to identify patients that can avail response from anti-EGFR therapy.

Prolapsed degenerating postpartum uterine fibroid: A case report.

Fibroid in pregnancy is common in clinical obstetric practice.There are conflicting reports in the literature on many so-called fibroid complications in pregnancy, and there are inadequate data on the optimum management strategy. We present a case of a pregnancy with large fibroid which was uneventful till delivery but ended in immediate postpartum complication of fibroid degenerating and prolapsing into vagina dealt with myomectomy at day 30 postnatal. Initially managed conservatively with intrauterine balloon tamponade and anaemia correction and discharged home. Later on myomectomy was performed on day 40 postop due to recurrent complaint of foul smelling vaginal discharge and fibroid protruding in vagina. This case illustrates a very rare complication of pregnancy with fibroid and the difficulties that are encountered in managing such cases.

Mismatch repair deficiency screening in colorectal carcinoma by a four-antibody immunohistochemical panel in Pakistani population and its correlation with histopathological parameters.

BACKGROUND: Microsatellite instability (MSI) operates as the second major pathway in the colorectal carcinogenesis. Although genetic testing remains the gold standard for the detection of MSI, the College of American Pathologists (CAP) recommends an initial immunohistochemical workup with a four-antibody panel (MLH1, PMS2, MSH2, and MSH6) to screen for a defective mismatch repair system. An increased trend towards young age colorectal carcinoma (CRC) has been noticed in our population over recent years; however, neither screening for MSI by immunohistochemistry (IHC)/genetic testing was done nor were its morphological features studied. We aimed to determine the frequency of mismatch repair deficiency (dMMR) by loss of IHC expression of the aforementioned enzymes in CRC patients and its correlatation with clinicopathologic parameters. METHODS: This was a retrospective study conducted at Liaquat National Hospital, Karachi, between 2012 and 2015. A total of 100 cases of CRC were included in the study that underwent surgical resection. IHC stains using antibodies MLH1, PMS2, MSH2, and MSH6 were performed by DAKO EnVision method on representative tissue blocks. The results were interpreted by senior histopathologists and correlated with clinico-pathological parameters. RESULTS: A total of 100 cases of CRC were studied that included 51 males and 49 females. Thirty-four percent (n = 34) of the patients showed loss of IHC staining for MMR markers. Combined loss of expression for MLH1/PMS2 were observed in 16% (n = 16) of the cases. Loss of MSH2/MSH6 were seen in 6% (n = 6) of the cases. Loss of expression for all markers were noted in 7% (n = 7) of the cases. There were 5% (n = 5) of the cases that showed isolated loss of MLH1 only. The tumors with dMMR status were significantly associated with right-sided location (p = 0.013), exhibited intra-tumoral lymphocytosis (p = 0.007), and lymphovascular invasion (p = 0.043). No significant association was seen with gender, age, tumor stage, grade, or other morphological features. CONCLUSION: The frequency of mismatch repair deficiency in CRC patients was found to be 34% in Pakistani population which warrants further genetic testing to exclude Lynch syndrome. Moreover, right-sided location and intra-tumoral lymphocyte count may be used to identify patients who may need further workup.

Increasing usage of rapid diagnostics for Dengue virus detection in Pakistan.

OBJECTIVE: To evaluate the trends in usage of dengue virus diagnostics in Pakistan. METHODS: This retrospective study was conducted at the Aga Khan University Hospital, Karachi, and comprised data for specimens tested for dengue virus from January 2012 to December 2015. Test for dengue virus ribonucleic acid by reverse transcription polymerase chain reaction, dengue virus antigen by immunochromatic assay and for human immunoglobulin M against dengue virus by enzyme-linked immunosorbent assay were reviewed. SPSS 17 was used for data analysis. RESULTS: Overall, 33,577 specimens tested for dengue virus. Of them, 11,995 (35.7%) were positive. among them, 1,039(8.66%) were reported in 2012; 5,791(48.28%) in 2013; 1,027(8.56%) in 2014; and 4,138(34.49%) in 2015. In 2012, 966(93%) of the positive samples were diagnosed by immunoglobulin M-based method and 73(7%) by non-structural protein-1 antigen. In 2013, 4,401(76%) samples were tested positive by immunoglobulin M, 1,332(23%) by antigen and 58(1%) by polymerase chain reaction. The trend continued in 2014, but in 2015, 2,111(51%) of all dengue positive tests were determined by antigen testing, 1,969(47.6%) by immunoglobulin M and 58(1.4%) by polymerase chain reaction. CONCLUSIONS: There was a shift in usage of direct virus identification for rapid diagnosis of dengue virus compared with host immunoglobulin M testing.

Antimicrobial susceptibility testing of Neisseria gonorrhoeae isolates in Pakistan by Etest compared to Calibrated Dichotomous Sensitivity and Clinical Laboratory Standards Institute disc diffusion techniques.

BACKGROUND: Accurate detection of Neisseria gonorrhoeae antimicrobial resistance is essential for appropriate management and prevention of spread of infection in the community. In this study Calibrated Dichotomous Sensitivity (CDS) and Clinical Laboratory Standards Institute (CLSI) disc diffusion methods were compared with minimum inhibitory concentration (MIC) by Etest in Neisseria gonorrhoeae isolates from Karachi, Pakistan. CDS and CLSI disc diffusion techniques, and Etest for ceftriaxone, penicillin G, spectinomycin and ciprofloxacin against 100 isolates from years 2012-2014 were performed. Due to lack of CLSI breakpoints for azithromycin, it was interpreted using cut-offs from British Society of Antimicrobial Chemotherapy (BSAC). Due to lack of low concentration tetracycline discs, tetracycline was tested with CLSI disc diffusion and Etest only. Comparisons were based on the identified susceptibility, intermediate susceptibility and resistance (SIR) categories using the different methods. Complete percent agreement was percentage agreement achieved when test and reference method had identical SIR-category. Essential percent agreement was percentage agreement when minor discrepancies were disregarded. RESULTS: There was 100 % and 99 % overall essential agreement and 50 % versus 23 % overall complete agreement by CDS and CLSI methods, respectively, with MICs for all tested antibiotics. Using either method, there was 100 % complete agreement for ceftriaxone and spectinomycin. There was 90 % versus 86 % complete agreement for ciprofloxacin, and 60 % and 75 % for penicillin using CDS and CLSI method, respectively. Essential agreement of 99 % and complete agreement of 62 % was found for tetracycline with CLSI method. There was 100 % essential and complete agreement by CDS, BSAC and Etest for azithromycin. CONCLUSION: No major errors with regard to identified SIR-categories were found for penicillin, ciprofloxacin, ceftriaxone and spectinomycin using CLSI and CDS methods. All isolates were susceptible to ceftriaxone and spectinomycin, and 99 % to azithromycin. In low-resource settings, both the CLSI and CDS disc diffusion techniques might be used for susceptibility testing of gonococcal isolates. However, these methods require considerable standardization and quality controls for adequate levels of reproducibility and correct interpretation to reflect appropriately the MIC values of the different antimicrobials. New, emerging, or rare resistance should be confirmed by MIC determination.