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CTRI Number  CTRI/2025/08/092654 [Registered on: 08/08/2025] Trial Registered Prospectively
Last Modified On: 07/08/2025
Post Graduate Thesis  Yes 
Type of Trial  Observational 
Type of Study   Cross Sectional Study 
Study Design  Other 
Public Title of Study   To Compare if microscopy, culture or PCR is a better method to diagnose eye disease caused by Acanthamoeba 
Scientific Title of Study   Optimizing diagnosis of Acanthamoeba Keratitis : A comparative study of Microscopy , Culture and PCR from a tertiary care centre of South India  
Trial Acronym  NIL 
Secondary IDs if Any  
Secondary ID  Identifier 
NIL  NIL 
 
Details of Principal Investigator or overall Trial Coordinator (multi-center study)  
Name  Dr Malavika Gopakumar 
Designation  Postgraduate 
Affiliation  Kasturba Medical College , Manipal 
Address  Department of Microbiology Centre for Basic Sciences Kasturba Medical College Madhav Nagar Manipal

Udupi
KARNATAKA
576104
India 
Phone  09048991953  
Fax    
Email  malavika1.kmcmpl2024@learner.manipal.edu  
 
Details of Contact Person
Scientific Query
 
Name  Dr Vinay Khanna 
Designation  Associate Professor 
Affiliation  Kasturba Medical College , Manipal 
Address  Department of Microbiology Centre for Basic Sciences Kasturba Medical College Madhav Nagar Manipal

Udupi
KARNATAKA
576104
India 
Phone  9964426900  
Fax    
Email  vinay.khanna@manipal.edu  
 
Details of Contact Person
Public Query
 
Name  Dr Vinay Khanna 
Designation  Associate Professor 
Affiliation  Kasturba Medical College , Manipal 
Address  Department of Microbiology Centre for Basic Sciences Kasturba Medical College Madhav Nagar Manipal

Udupi
KARNATAKA
576104
India 
Phone  9964426900  
Fax    
Email  vinay.khanna@manipal.edu  
 
Source of Monetary or Material Support  
Institutional PG Thesis Fund , Kasturba Medical College , MAHE, Madhav Nagar, Manipal, Udupi Karnataka PINCODE 576104 
 
Primary Sponsor  
Name  Kasturba Medical College , Manipal 
Address  Kasturba Medical College , Madhav Nagar Manipal Udupi Pincode 576104  
Type of Sponsor  Private medical college 
 
Details of Secondary Sponsor  
Name  Address 
NIL  NIL 
 
Countries of Recruitment     India  
Sites of Study  
No of Sites = 1  
Name of Principal Investigator  Name of Site  Site Address  Phone/Fax/Email 
Dr Malavika Gopakumar  Kasturba Medical College , Manipal  Department of Microbiology, Centre for Basic Sciences Kasturba Medical College Madhav Nagar,Eshwar Nagar Manipal , Udupi Kasturba Medical College Madhav Nagar, Manipal Karnataka PINCODE 576104
Udupi
KARNATAKA 
09048991953

malavika1.kmcmpl2024@learner.manipal.edu 
 
Details of Ethics Committee  
No of Ethics Committees= 1  
Name of Committee  Approval Status 
Kasturba Medical College and Kasturba Hospital  Approved 
 
Regulatory Clearance Status from DCGI  
Status 
Not Applicable 
 
Health Condition / Problems Studied  
Health Type  Condition 
Healthy Human Volunteers  any diseases in eye other than acanthamoeba or infectious keratitis 
Patients  (1) ICD-10 Condition: H168||Other keratitis, (2) ICD-10 Condition: H160||Corneal ulcer,  
 
Intervention / Comparator Agent  
Type  Name  Details 
Intervention  Nil  Nil 
Comparator Agent  Nil  Nil 
 
Inclusion Criteria  
Age From  1.00 Day(s)
Age To  95.00 Year(s)
Gender  Both 
Details  Patients of any age and gender presenting with symptoms suggestive of Acanthamoeba
keratitis (pain, redness, photophobia, blurred vision). 
 
ExclusionCriteria 
Details  1)Patients with a history of recent ocular surgery or trauma that may introduce non-Acanthamoeba pathogens.
2)Patients already receiving treatment for confirmed Acanthamoeba keratitis or any other severe
ocular infection. 
 
Method of Generating Random Sequence   Not Applicable 
Method of Concealment   Not Applicable 
Blinding/Masking   Not Applicable 
Primary Outcome  
Outcome  TimePoints 
1)To find out diagnostic accuracy in terms of Sensitivity , Specificity, PPV, NPV and accuracy for
microscopy , culture ,c-PCR for Acanthamoeba Keratitis

2)To find out proportion of Acanthamoeba Keratitis cases among clinically suspected infectious keratitis 
Baseline 
 
Secondary Outcome  
Outcome  TimePoints 
Identification of risk factors associated with Acanthamoeba Keratitis in terms of delayed diagnosis , contact lens use,immunosuppression etc  Baseline 
 
Target Sample Size   Total Sample Size="180"
Sample Size from India="180" 
Final Enrollment numbers achieved (Total)= "Applicable only for Completed/Terminated trials"
Final Enrollment numbers achieved (India)="Applicable only for Completed/Terminated trials" 
Phase of Trial   N/A 
Date of First Enrollment (India)   20/08/2025 
Date of Study Completion (India) Applicable only for Completed/Terminated trials 
Date of First Enrollment (Global)  Date Missing 
Date of Study Completion (Global) Applicable only for Completed/Terminated trials 
Estimated Duration of Trial   Years="1"
Months="6"
Days="0" 
Recruitment Status of Trial (Global)   Not Applicable 
Recruitment Status of Trial (India)  Not Yet Recruiting 
Publication Details   N/A 
Individual Participant Data (IPD) Sharing Statement

Will individual participant data (IPD) be shared publicly (including data dictionaries)?  

Response - NO
Brief Summary   Acanthamoeba, a genus of free-living amoebae, inhabits diverse environments such as soil, freshwater, air, sewage, chlorinated water, medical equipment, and contact lens cases. These amoebae exist as trophozoites, actively feeding and proliferating under favorable conditions, but transform into highly resistant cysts under stress. While primarily bacterivorous, certain species, including Acanthamoeba castellanii, A. polyphaga, and A. hatchetti, exhibit pathogenic potential, causing keratitis and granulomatous amoebic encephalitis (GAE), particularly in immunocompromised individuals. GAE occurs when amoebae enter through wounds and disseminate to the brain. Acanthamoeba has also been detected on human skin, in the respiratory and gastrointestinal tracts, and in other hosts, highlighting its ecological adaptability and clinical significance.

Acanthamoeba castellanii, A. polyphaga, and A. hatchetti are among the primary species implicated in Acanthamoeba keratitis (AK), a severe corneal infection that predominantly affects contact lens users due to improper lens hygiene. Hydrogen peroxide-based disinfectants effectively eliminate Acanthamoeba, whereas multipurpose solutions are largely ineffective. AK is often misdiagnosed due to its nonspecific symptoms, delaying treatment and increasing the risk of vision loss. First identified in 1973, its incidence rose in the 1980s with the widespread use of contact lenses, which account for 80–93% of cases globally. In contrast, in rural areas of developing countries, corneal trauma from vegetative matter is the primary risk factor, highlighting distinct epidemiological patterns. 

The incidence of Acanthamoeba keratitis (AK) has risen significantly, with cases in the United States increasing tenfold from 2 million per year in 1988 to 20 million in 2003 among contact lens wearers. A study in South India analyzing over 38,000 infectious keratitis cases attributed 0.9% to AK. Despite this growing burden, global prevalence data remain limited, with most knowledge derived from localized studies and case reports, particularly those linked to contact lens use. 

With the increasing incidence of Acanthamoeba keratitis (AK), along with diagnostic challenges and limited treatment options, further research is crucial. This study compares two diagnostic methods— Polymerase Chain Reaction (PCR) and culture isolation—for detecting Acanthamoeba species in AK cases. PCR is a highly sensitive technique that can identify even small amounts of Acanthamoeba DNA, making it particularly useful for early diagnosis. In contrast, culture isolation allows for species identification and provides biological insights by growing the amoeba from clinical samples. This study aims to evaluate the accuracy, effectiveness, and practical use of PCR versus culture isolation in AK diagnosis. Identifying the most reliable method will improve diagnostic protocols, enhance patient outcomes, and support public health strategies for better prevention.
 
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